Heterocyclic compounds and methods of use

ABSTRACT

Heterocyclic compounds derived from benzotriazine, triazines, triazoles and oxadiazoles are disclosed. The methods of synthesis and of use of such heterocyclic compounds are also provided.

CROSS-REFERENCE TO RELATED APPLICATIONS

This application is a continuation application of U.S. patentapplication Ser. No. 12/684,312 filed Jan. 8, 2010, which is acontinuation application of U.S. patent application Ser. No. 11/212,064filed Aug. 24, 2005, which claims the benefit of priority under 35U.S.C. §119(e) of U.S. Provisional Patent Application Nos. 60/604,298filed Aug. 25, 2004, and 60/696,168 filed Jul. 1, 2005, the disclosuresof which are incorporated herein by reference in their entirety.

FIELD OF THE INVENTION

The present invention relates generally to the use of compounds to treata variety of disorders, diseases and pathologic conditions and morespecifically to the use of various heterocyclic compounds fortherapeutic purposes.

BACKGROUND

Kinases are a large family of cellular proteins involved in signaltransduction of cascades which control cell growth and death, survival,migration, differentiation, gene expression, metabolism, proteinsynthesis and cell cycle regulation. A common mechanism by which thesesignals are transmitted is reversible phosphorylation, which inducesconformational changes is these enzymes and alters their structure andfunction. The entire kinase genome discovered so far incorporates over500 individual proteins and their isoforms. Different branches of thisgenomic tree have been characterized into groups specific forphosphorylating either serine/threonine residues or tyrosines. Somekinases exhibit dual specificity, capable of substrate phosphorylationof tyrosine as well as serine/threonines. Further differentiation can bemade in terms of their location in cells. Transmembrane receptor proteinkinases exhibit an extracellular domain, capable of ligand binding.These ligand binding mechanisms trigger activation of the kinasecatalytic domain which initiates a cascade of signals that controlsintracellular functions. Examples of a receptor protein kinase aregrowth factors such as EGF, FGF, PDGF and IGF. Nonreceptor proteinkinases can be found in many compartments of a cell from inner-cellsurface membranes to the cell nucleus. One example of a nonreceptorprotein kinase is the mitogen activated protein kinase (MAPK) whichregulates a pathway, which is important in cell signaling initiated onthe exterior cell surfaces via growth factors, for example, VEGF, orhormones, and extending to the cell nucleus by activating transcriptionfactors. These nuclear factors in turn control gene expression in theregulation of cell cycle progression and ultimately cell proliferation,and differentiation.

The MAPK cell signaling pathway is important for drug targeting as thispath impinges on nearly all functional hallmarks of cancer cells such asimmortalization, growth factor independent proliferation, insensitivityto growth inhibitory signals, metastasis, blood vessel attraction,evasion of apoptosis, and other functional hallmarks. Inappropriateactivation though mutation of this molecule is associated with nearly30% of all human cancers. In general, the inhibition of disregulatedkinases such as Ras, PI3K and Raf is an important approach to discovernovel treatments for cancer and other diseases. One approach is thediscovery of small molecules capable of binding either to the kinasecatalytic domain or a regulatory domain in order to modulate thefunction of protein kinases. Important in this respect is to discovermolecules which inhibit a specific signaling path with a high degree ofselectivity and a potency within a practical therapeutic window. Whilesignificant progress has been made in developing various compounds forthe treatment of cancer and inflammatory diseases, there remains a needfor specific chemical structures capable of modulating protein kinases,whose disregulated function has been implicated in these diseases.

SUMMARY OF THE INVENTION

The present invention provides compounds which affect the MAPK pathway.The compounds of the invention are useful as pharmaceuticalcompositions, for example where modulation of the MAPK pathway isindicated for the treatment of various human diseases, such as cancer.

According to one embodiment of the invention, compounds having thestructure (A) are provided, or an N-oxide, N,N′-dioxide,N,N′,N″-trioxide, or a pharmaceutically acceptable salt thereof:

wherein Y can be absent or can be one of the following moieties:

According to another embodiment of the invention, compounds having thestructure (B) are provided, or an N-oxide, N,N′-dioxide,N,N′,N″-trioxide, or a pharmaceutically acceptable salt thereof:

wherein X can be absent or can be NH, and Y can be absent or can be oneof the following moieties:

In compounds having structure (B), each of Z₁, Z₂ and Z₃ can be,independently, N, N═CH, CH, O, S or N—R⁴, wherein R⁴ is hydrogen orlower alkyl, with the further proviso that at least one of Z₁, Z₂ and Z₃is not CH.

In compounds having structure (A), the substitutent R₁ can be an aryl, asubstituted aryl, a heterocycle, a heteroaryl, a substitutedheterocycle, and a substituted heteroaryl. For example, R₁ can be one ofC₆-C₁₂ aryl; C₃-C₁₂ heteroaryl having 1-3 heteroatoms such as N, S andO; substituted C₃-C₁₀ cycloalkyl having 0-3 heteroatoms such as N, S,and O; substituted C₆-C₁₂ aryl; substituted C₃-C₁₂ heteroaryl having 1-3heteroatoms such as N, S and O; C₇-C₂₄ aralkyl; C₇-C₂₄ alkylaryl;substituted C₇-C₂₄ aralkyl; and substituted C₇-C₂₄ alkaryl.

In compounds having structure (B), the substitutent R₁ can be,independently of the substitutent R₁ present in the structure (A), anunsubstituted or a substituted C₃-C₁₂ heteroaryl having 1-3 heteroatomssuch as N, S or O. The substituent R₁ that can be present in compoundshaving structure (B), can include a substituted pyridyl group. Thesubstituents in the substituted pyridyl group can include an amidomoiety, an aminoalkyl group (e.g., aminomethyl), or a carboxyl group, ora carboxylate group. The amido moiety attached to the pyridyl group canbe in turn also substituted by attaching to the nitrogen in the amidomoiety a substitutent selected from an alkyl (e.g., methyl), analkylaminoalkyl (e.g., diethylamino alkyl), a pyridyl, an alkylpyrrolidine, an alkyl morpholine, and an alkyl piperazine groups.

Some examples of the substitutent R₁ that can be present in compoundshaving either structure (A) or structure (B), can be selected from oneof the following moieties:

where n can be an integer selected from a group consisting of 0, 1, 2,and 3.

In compounds having structure (A) or (B), R₂ can be, independently, anyone of hydrogen, halogen, C₁-C₁₈ alkyl (e.g., methyl), —OH, —NO₂, —CN,C₁-C₁₈ alkoxy (e.g., methoxy), —NHSO₂R⁵, —SO₂NHR⁵, —NHCOR⁵, —NH₂,—NR⁵R⁶, —S(O)R⁵, —S(O)₂R⁵, —CO₂R⁵, —CONR⁵R⁶, and where R⁵ and R⁶ areindependently selected from hydrogen, a C₁-C₁₂ alkyl and a substitutedC₁-C₁₂ alkyl.

In compounds having structure (A), the substituent R₃ can be an aryl, asubstituted aryl, a heterocycle, a heteroaryl, a substitutedheterocycle, and a substituted heteroaryl. For example, R₃ can be one ofC₆-C₁₂ aryl; C₃-C₁₂ heteroaryl having 1-3 heteroatoms such as N, S andO; substituted C₃-C₁₀ cycloalkyl having 0-3 heteroatoms such as N, S,and O; substituted C₆-C₁₂ aryl; substituted C₃-C₁₂ heteroaryl having 1-3heteroatoms such as N, S and O; C₇-C₂₄ aralkyl; C₇-C₂₄ alkylaryl;substituted C₇-C₂₄ aralkyl; and substituted C₇-C₂₄ alkaryl.

In compounds having structure (B), the substitutent R₃ can be,independently of the substitutent R₃ present in the structure (A),hydrogen, a C₁-C₁₈ alkyl, a substituted C₁-C₁₈ alkyl, a C₁-C₁₂cycloalkyl, a substituted C₁-C₁₂ cycloalkyl, a substituted C₃-C₁₀cycloalkyl having 0-3 heteroatoms such as N, S, or O, an aryl such as aC₆-C₁₂ aryl, a substituted aryl such as a substituted C₆-C₁₂ aryl, aheterocycle, a substituted heterocycle, a heteroaryl such as a C₃-C₁₂heteroaryl having 1-3 heteroatoms such as N, S or O, a substitutedheteroaryl such as substituted C₃-C₁₂ heteroaryl having 1-3 heteroatomssuch as N, S or O, a C₇-C₂₄ aralkyl, a substituted C₇-C₂₄ aralkyl, aC₇-C₂₄ alkylaryl, and a substituted C₇-C₂₄ alkaryl. Some particularexamples of the substituent R₃ than can be used include tert-butylphenyl, trifluoromethoxyphenyl, methoxyphenyl, dimethylaminophenyl,aminophenyl, trifluoroethoxyphenyl, trifluoromethoxychlorophenyl,trifluoromethoxybromophenyl, trifluoroethoxychlorophenyl, chlorophenyl,dichlorophenyl, trifluoromethyl phenyl, trifluoromethylchlorophenyl,chlorotoluyl, N-phenylacetamide, N,N-alkyl-benzamide, isopropoxyphenyl,alkoxyphenyl, dialkoxyphenyl, or acetylphenyl.

To summarize, some examples of the substitutent R₃ that can be presentin compounds having either structure (A) or structure (B), can beselected from one of the following moieties:

According to another embodiment of the invention, compounds that arederivatives of benzotriazine are provided, the compounds including abenzotriazine moiety having at least a first substituent attached to thebenzene ring of benzotriazine and a second substituent attached to thetriazine ring of the benzotriazine, where the first substituent includesa substituted pyridyl group, and the second substituent includes asecondary amino group, a substituted amide group, or a substitutedsulfonylamino group.

According to yet another embodiment of the invention, compoundsincluding a benzene-derived moiety bridged to a heterocyclic moiety areprovided, where the benzene-derived moiety includes a molecule ofbenzene substituted with either a sulfonyl group or a pyridyl groupconnected to the benzene molecule via an oxygen link, and theheterocyclic moiety includes triazole, oxadiazole, oxazole, pyrazol,imidazole, thiadiazole and triazine.

According to yet another embodiment, articles of manufacture areprovided, the articles including packaging material and a pharmaceuticalcomposition contained within the packaging material, wherein thepackaging material includes a label which indicates that thepharmaceutical composition can be used for treatment of disordersassociated with cancer. The pharmaceutical composition can include atleast one compound set forth in Structures (A) and (B) or anycombination thereof.

According to another embodiment, a method for treating a disorderincluding administering to a subject in need thereof an effective amountof a compound, wherein the compound is set forth in Structures (A) and(B) or any combination thereof.

In one aspect, the disorder is cancer, eye disease, inflammation,psoriasis, or a viral infection, for example. More particularly, thecancer is an alimentary/gastrointestinal tract cancer, colon cancer,liver cancer, skin cancer, breast cancer, ovarian cancer, prostatecancer, lymphoma, leukemia, kidney cancer, lung cancer, muscle cancer,bone cancer, bladder cancer or brain cancer.

According to yet another embodiment, a pharmaceutical composition isprovided, at least one compound set forth in structures (A) and (B) orany combination thereof, in a pharmaceutically acceptable carrier.

BRIEF DESCRIPTION OF THE DRAWINGS

FIG. 1 shows the results of the Raf1 direct assay of invention compounds

FIG. 2 shows the results of XTT cell viability assay of inventioncompounds

DETAILED DESCRIPTION OF THE INVENTION

The present invention is directed to heterocyclic compounds, such asheterocyclic compounds derived from benzotriazine, triazines, triazoles,oxadiazoles, imidazoles and thiadiazole and to use of the heterocycliccompounds for therapeutic purposes.

The following terminology and definitions apply as used in the presentapplication, generally in conformity with the terminology recommended bythe International Union of Pure and Applied Chemistry (IUPAC).

The term “heterocyclic,” when used to describe an aromatic ring, meansthat the aromatic ring contains at least one heteroatom. Theabbreviation “Het” is sometimes used to signify a heterocyclicstructure.

The term “heteroatom” is defined to include any atom other than carbon,for example, N, O, or S.

The term “aromatic” or “aryl” is defined to include a cyclicallyconjugated molecular entity with a stability, due to delocalization,significantly greater than that of a hypothetical localized structure,such as the Kekulé structure.

The term “heterocyclic,” when not used to describe an aromatic ring, isdefined to include cyclic (i.e., ring-containing) groups other thanaromatic groups, the cyclic group being formed by between 3 and about 14carbon atoms and at least one heteroatom described above.

The term “substituted heterocyclic” is defined to include both aromaticand non-aromatic structures to heterocyclic groups further bearing oneor more substituents described above.

The term “alkyl” is defined to include a monovalent straight or branchedchain hydrocarbon group having from one to about 12 carbon atoms, forexample, methyl, ethyl, n-propyl, iso-propyl, n-butyl, iso-butyl,tert-butyl, n-pentyl (also known as n-amyl), n-hexyl, and the like. Theabbreviations “Me” and “Et” stand for the methyl and ethyl groups,respectively.

The term “aralkyl” refers to an aryl or heteroaryl group, as definedherein, attached through a C1-C6 alkyl linker. Examples of “aralkyl”include, but are not limited to, benzyl, phenylpropyl, 2-pyridylmethyl,3-isoxazolylmethyl, 2-imidazolylethyl.

The term “methoxy” is defined as the group —OCH₃.

The term “halogen” is defined to include an atom of fluorine, chlorine,bromine or iodine.

The term “carboxyl” or “carboxyl group” is defined as an acid moietyhaving the structure —COOH.

The term “amino” or “amino group” is defined to include moieties —NRR′,where each of R and R′ is hydrogen (“primary amino”), or one of them isan organic radical (“secondary amino”), or each is an organic radical(“tertiary amino”).

The term “aminoalkyl” or “aminoalkyl group” is defined to includemoieties —R—N(R′R″), wherein R is an organic radical and each of R′ andR″ is hydrogen or an organic radical. If at least one of R′ and R″ is anorganic radical, the moiety is defined as “alkylaminoalkyl” or“alkylaminoalkyl group.”

The term “sulfonyl” or “sulfonyl group” is defined to include moietiesthat comprise structure (S), in which R is an organic radical:

The term “sulfonylamino” or “sulfonylamino group” is defined to includemoieties that comprise structure (SA), in which R is an organic radical:

The term “amide,” or “amido,” or “amide group,” or “amido group” isdefined to include moieties containing at least one acyl group >C═Oattached to nitrogen. The term “substituted amide” is defined to includemoieties containing a structure RNH—CO—, in which R is an organicradical.

The term “phenyl” is defined to include moieties having structure (Ph):

The term “toluoyl” is defined to include moieties having structure (Tl):

The term “heteroaryl” is defined to include aromatic rings, where thering structure is formed by between 3 and about 14 carbon atoms and byat least one heteroatom described above, and the term “substitutedheteroaryl” refers to heteroaryl groups further bearing one or moresubstituents described above.

The term “triazine” is defined to include moieties containing thearomatic 6-member heterocycle having three atoms of nitrogen in thering. Two examples of such heterocycle are shown as the followingstructures (Tr):

The term “benzotriazine” is defined to include moieties containing aheterocyclic structure in which the triazine ring is fused with thebenzene ring, as shown by structure (BTr):

The terms “N-oxide,” “N,N′-dioxide,” and “N,N′,N″-trioxide” are definedto include nitrogen-containing heterocyclic moieties in which at leastone nitrogen atom is associated with oxygen to form the structures N→O.The heterocyclic moiety can be any nitrogen-containing heterocycle, forexample, benzotriazine, triazine, pyridine, pyrimidine, etc. Where theheterocyclic structure is benzotriazine, for example, some N-oxides ordioxides can be described as the following structures:

The term “pteridine” is defined to include moieties containing aheterocyclic structure having two fused 6-member rings, each ringcontaining two atoms of nitrogen, as shown by structure (PTr):

The term “pyridazine” is defined to include moieties containing thearomatic 6-member heterocycle having two atoms of nitrogen in the ringin ortho position, as shown by the structure (PAz):

The term “pyrimidine” is defined to include moieties containing thearomatic 6-member heterocycle having two atoms of nitrogen in the ringin meta position, as shown by the structure (PRm):

The term “thiadiazole” is defined to include moieties containing thearomatic 5-member thiophene-based heterocycle, having two atoms ofnitrogen and one atom of sulfur, as shown by the structure (TDa):

The term “pyridyl” is defined to include moieties containing a radicalderived from pyridine. One structure of pyridyl is shown as thestructure (Py):

The term “alkyl pyrrolidine” is defined to include moieties containing aradical derived from pyrrolidine (a 5-member saturated heterocyclehaving one nitrogen atom), where an alkylene group R is attached to thenitrogen atom of the pyrrolidine ring. One structure of alkylpyrrolidine is shown as the structure (APy):

The term “alkyl morpholine” is defined to include moieties containing aradical derived from morholine, (a 6-member saturated heterocycle havingone nitrogen atom and one oxygen atom), where an alkylene group R isattached to the nitrogen atom of the morpholine ring. One structure ofalkyl morpholine is shown as the structure (AMr):

The term “alkyl piperazine” is defined to include moieties containing aradical derived from piperazine (a 6-member saturated heterocycle havingtwo nitrogen atoms), where an alkylene group R is attached to onenitrogen atom of the piperazine ring. One structure of alkyl piperazineis shown as the structure (APi):

The term “isoxazole” is defined to include moieties containing thearomatic 5-member heterocycle, having one atoms of nitrogen and one atomof oxygen, as shown by the structure (ISo):

The term “hydrophobic” is defined as a group or structure free ofstrongly polar groups such as —OH, —COOH, —NH₂, —NH—CO—, halogens, orthe like.

The term “kinase” is defined to include any enzyme that catalyzes theaddition of phosphate groups to a protein residue; for example, serineand threonine kinases catalyze the addition of phosphate groups toserine and threonine residues.

The term “therapeutically effective amount” is defined as the amount ofthe compound or pharmaceutical composition that will elicit thebiological or medical response of a tissue, system, animal or human thatis being sought by the researcher, veterinarian, medical doctor or otherclinician, e.g., restoration or maintenance of vasculostasis orprevention of the compromise or loss or vasculostasis; reduction oftumor burden; reduction of morbidity and/or mortality.

The term “pharmaceutically acceptable” is defined as a carrier, whetherdiluent or excipient, that is compatible with the other ingredients ofthe formulation and not deleterious to the recipient thereof.

The terms “administration of a compound” or “administering a compound”is defined to include an act of providing a compound of the invention orpharmaceutical composition to the subject in need of treatment.

According to embodiments of the present invention, two types ofheterocyclic compounds are provided for treatment of various diseases,disorders, and pathologies, including cancer. The heterocyclic compoundsof the invention can inhibit the activity of a kinase, such as anykinase in the MAPK signaling pathway.

According to an embodiment of the invention, a first type of compoundsis provided for treatment of various diseases, disorders, andpathologies, including cancer. The first type of compounds can includederivatives of benzotriazine. The derivatives of benzotriazine that canbe used can comprise the compounds that include a benzotriazine moietyhaving at least one substituent attached to the benzene ring ofbenzotriazine and a at least one substituent attached to the triazinering of the benzotriazine, and an N-oxide, N,N′-dioxide,N,N′,N″-trioxide, or pharmaceutically acceptable salts thereof.

A substituent attached to the benzene ring of benzotriazine can includea substituted pyridyl group. The substituents in the substituted pyridylgroup can include an amido moiety, an aminoalkyl group (e.g.,aminomethyl), or a carboxyl group, or a carboxylate group. The amidomoiety attached to the pyridyl group can be in turn also substituted byattaching to the nitrogen in the amido moiety a substituent selectedfrom an alkyl (e.g., methyl), an alkylaminoalkyl (e.g., diethylaminoalkyl), a pyridyl, an alkyl pyrrolidine, an alkyl morpholine, and analkyl piperazine groups.

Optionally, the benzene ring of the compounds of the first type cancontain a second substituent located in any available position of thering, for example, methyl, halogen or methoxy. Some particular examplesof benzene-ring containing moieties that can be used include tert-butylphenyl, trifluoromethoxyphenyl, methoxyphenyl, dimethylamino,dimethylaminophenyl, aminophenyl, trifluoroethoxyphenyl,trifluoromethoxychlorophenyl, trifluoromethoxybromophenyl,trifluoroethoxychlorophenyl, chlorophenyl, dichlorophenyl,trifluoromethyl phenyl, trifluoromethylchloro phenyl, chlorotoluyl,N-phenylacetamide, N,N-alkyl-benzamide, isopropoxyphenyl, alkoxyphenyl,dialkoxyphenyl, acetylphenyl.

A substituent attached to the triazine ring of benzotriazine in thecompounds of the first type can include a secondary amino group, asubstituted amide group, or a substituted sulfonylamino group; each ofthese groups can further contain a moiety derived from benzene,thiophene, or isoxazole. If the substituent attached to the triazinering of benzotriazine in the compounds of the first type is a secondaryamino group, the moiety derived from benzene, thiophene, or isoxazolecan be attached to the nitrogen of the secondary amino group. If thesubstituent in the triazine ring is a substituted amide group or thesubstituted sulfonylamino group, the moiety derived from benzene,thiophene, or isoxazole can be attached via the acyl group or thesulfonyl group, respectively. Moieties derived from benzene, thiophene,or isoxazole can further include alkyls, e.g., t-butyl phenyl,chlorophenyl, dichlorophenyl, trifluoromethyl phenyl,trifluoromethylchloro phenyl; and chlorotoluyl.

Compounds of the first type can be described as compounds having thegeneral structure (A), and an N-oxide, N,N′-dioxide, N,N′,N″-trioxide,or a pharmaceutically acceptable salt thereof. The general structure (A)is as follows:

wherein Y can be absent or can be one of the following moieties:

In structure (A), R₁ can be an aryl, a substituted aryl, a heterocycle,a heteroaryl, a substituted heterocycle, and a substituted heteroaryl,such as C₆-C₁₂ aryl; C₃-C₁₂ heteroaryl having 1-3 heteroatoms such as N,S and O; substituted C₃-C₁₀ cycloalkyl having 0-3 heteroatoms such as N,S, and O; substituted C₆-C₁₂ aryl; substituted C₃-C₁₂ heteroaryl having1-3 heteroatoms such as N, S and O; C₇-C₂₄ aralkyl; C₇-C₂₄ alkylaryl;substituted C₇-C₂₄ aralkyl; and substituted C₇-C₂₄ alkaryl. Inparticular, R₁ can be any one of the following moieties:

where n can be an integer selected from a group consisting of 0, 1, 2,and 3.

In structure (A), the substituent R₂ can be any one of hydrogen,halogen, C₁-C₁₈ alkyl (e.g., methyl), —OH, —NO₂, —CN, C₁-C₁₈ alkoxy(e.g., methoxy), —NHSO₂R⁵, —SO₂NHR⁵, —NHCOR⁵, —NH₂, —NR⁵R⁶, —S(O)R⁵,—S(O)₂R⁵, —CO₂R⁵, —CONR⁵R⁶, and where R⁵ and R⁶ are independentlyselected from hydrogen, a C₁-C₁₂ alkyl and a substituted C₁-C₁₂ alkyl.

In structure (A), group R₃ can be an aryl, a substituted aryl, aheterocycle, a heteroaryl, a substituted heterocycle, and a substitutedheteroaryl. For example, R₃ can be one of C₆-C₁₂ aryl; C₃-C₁₂ heteroarylhaving 1-3 heteroatoms such as N, S and O; substituted C₃-C₁₀ cycloalkylhaving 0-3 heteroatoms such as N, S, and O; substituted C₆-C₁₂, aryl;substituted C₃-C₁₂ heteroaryl having 1-3 heteroatoms such as N, S and O;C₇-C₂₄ aralkyl; C₇-C₂₄ alkylaryl; substituted C₇-C₂₄ aralkyl; andsubstituted C₇-C₂₄ alkaryl. In particular, the substitutent R₃ can beone of the following:

Some general examples of compounds described by the general structure(A) include a compound having the general structure (I), a compoundhaving the general structure (II), a compound having the generalstructure (III) or a compound having the general structure (IV):

Some non-limiting examples of particular compounds described by thegeneral structure (A) that can be used include compounds having formulae(1)-(33):

The benzotriazine derivatives described above and illustrated by thegeneral structure (A) can be prepared as shown by Scheme I:

To prepare the intermediate A, where R₁ is, for example, 2-pyridinecarboxamide, the synthetic route shown by Scheme II can be used:

As seen from Scheme II, the synthesis of the intermediate A requiresusing 4-chloro-2-pyridinecarboxamide 4, which can be separatelypreliminarily synthesized as shown by Scheme III:

The benzotriazine derivatives described in the formula (IV) andillustrated by the general structure (A) can be prepared as shown byScheme IV:

According to an embodiment of the invention, a second type of compoundsis provided for treatment of various diseases, disorders, andpathologies, including cancer. The second type of compounds can includea benzene derived moiety bridged to a heterocyclic moiety, orpharmaceutically acceptable salts thereof. The bridge between thebenzene-derived moiety and the heterocyclic moiety can include a singlebond or a nitrogen atom. If the heterocyclic moiety contains at leastone nitrogen, the second type of compounds can be an N-oxide, orN,N′-dioxide, or N,N′,N″-trioxide.

Whether the compound can be an N-oxide, or N,N′-dioxide, orN,N′,N″-trioxide, depends on the number of nitrogen atoms contained inthe heterocyclic moiety. For example, if the heterocyclic moiety hasonly one nitrogen, the second type of compounds can be an N-oxide. Ifthe heterocyclic moiety has two atoms of nitrogen, the second type ofcompounds can be an N-oxide or N,N′-dioxide. If the heterocyclic moietyhas three atoms of nitrogen, the second type of compounds can be any ofan N-oxide, an N,N′-dioxide, an N,N′,N″-trioxide.

The benzene-derived moiety can include a substituent such as a pyridylgroup connected to the benzene molecule via an oxygen link, or asulfonyl group. The pyridyl group connected to the benzene molecule canbe further substituted. The substituents in the pyridyl group caninclude the same moieties as described above for the first type ofcompounds of the present invention. The sulfonyl group connected to thebenzene molecule can be also further substituted. The substituents inthe sulfonyl group can include the substituted pyridyl group describedabove for the first type of compounds of the present invention.

Optionally, the benzene-derived moiety of the compounds of the secondtype can contain a second substituent, e.g., methyl, halogen or methoxy,which can be located in any position of the benzene ring. Some exemplarybenzene-derived moieties that can be included in the second type ofcompounds can include tert-butyl phenyl, trifluoromethoxyphenyl,methoxyphenyl, dimethylamino, dimethylaminophenyl, aminophenyl,trifluoroethoxyphenyl, trifluoromethoxychlorophenyl,trifluoromethoxybromophenyl, trifluoroethoxychlorophenyl, chlorophenyl,dichlorophenyl, trifluoromethyl phenyl, trifluoromethylchloro phenyl,chlorotoluyl, N-phenylacetamide, N,N-alkyl-benzamide, isopropoxyphenyl,alkoxyphenyl, dialkoxyphenyl, acetylphenyl.

The compounds of the second type include heterocyclic compounds havingthe general structure (B), or an N-oxide, or N,N′-dioxide,N,N′,N″-trioxide, or a pharmaceutically acceptable salt thereof, and caninhibit the activity of a kinase, such as any kinase in the MAPKsignaling pathway. The general structure (B) can be represented asfollows:

In structure (B), each of Z₁, Z₂ and Z₃ can be, independently, N, CH,N═CH, O, S or N—R⁴, wherein R⁴ is hydrogen or lower alkyl, with thefurther proviso that at least one of Z₁, Z₂ and Z₃ is not CH; X can beabsent or be NH; Y can be absent or can be one of the followingmoieties:

Further, in structure (B), the substitutents R₁, R₂, and R₃ can be asfollows:

R₁ can be an unsubstituted or a substituted C₃-C₁₂ heteroaryl having 1-3heteroatoms such as N, S or O;

R₂ can be any one of hydrogen, halogen, C₁-C₁₈ alkyl (e.g., methyl),—OH, —NO₂, —CN, C₁-C₁₈ alkoxy (e.g., methoxy), —NHSO₂R⁵, —SO₂NHR⁵,—NHCOR⁵, —NH₂, —NR⁵R⁶, —S(O)R⁵, —S(O)₂R⁵, —CO₂R⁵, —CONR⁵R⁶, and where R⁵and R⁶ are independently selected from hydrogen, a C₁-C₁₈ alkyl and asubstituted C₁-C₁₂ alkyl; and

R₃ can be hydrogen, a C₁-C₁₈ alkyl, a substituted C₁-C₁₂ alkyl, a C₁-C₁₂cycloalkyl, a substituted C₁-C₁₂ cycloalkyl, a substituted C₃-C₁₀cycloalkyl having 0-3 heteroatoms such as N, S, or O, an aryl such as aC₆-C₁₂ aryl, a substituted aryl such as a substituted C₆-C₁₂ aryl, aheterocycle, a substituted heterocycle, a heteroaryl such as a C₃-C₁₂heteroaryl having 1-3 heteroatoms such as N, S or O, a substitutedheteroaryl such as substituted C₃-C₁₂ heteroaryl having 1-3 heteroatomssuch as N, S or O, a C₇-C₂₄ aralkyl, a substituted C₇-C₂₄ aralkyl, aC₇-C₂₄ alkylaryl, and a substituted C₇-C₂₄ alkaryl.

The substituent R₁ can include a substituted pyridyl or a substitutedpyrimidyl group. The substituents in the substituted pyridyl orsubstituted pyrimidyl group can include an amido moiety, an aminoalkylgroup (e.g., aminomethyl), or a carboxyl group, or a carboxylate group.The amido moiety attached to the pyridyl/pyrimidyl group can be in turnalso substituted by attaching to the nitrogen in the amido moiety asubstitutent selected from an alkyl (e.g., methyl), an alkylaminoalkyl(e.g., diethylamino alkyl), a pyridyl, an alkyl pyrrolidine, an alkylmorpholine, and an alkyl piperazine groups.

Particular, non-limiting examples of R₁ that can be used in compoundshaving the structure (B) include any of the following moieties:

where n can be an integer selected from a group consisting of 0, 2, and3.

Some particular non-limiting examples of the substituent R₃ that can beused in compounds having the structure (B) include tert-butyl phenyl,trifluoromethoxyphenyl, methoxyphenyl, dimethylaminophenyl, aminophenyl,trifluoroethoxyphenyl, trifluoromethoxychlorophenyl,trifluoromethoxybromophenyl, trifluoroethoxychlorophenyl, chlorophenyl,dichlorophenyl, trifluoromethyl phenyl, trifluoromethylchlorophenyl,chlorotoluyl, N-phenylacetamide, N,N-alkyl-benzamide, isopropoxyphenyl,alkoxyphenyl, dialkoxyphenyl, and acetylphenyl. These and yet othermoieties that can be used as the substituent R₃ can be illustrated asfollows:

where n can be an integer selected from a group consisting of 0, 1, 2,and 3, and R′ is hydrogen, a C₁-C₁₈ alkyl, or a substituted C₁-C₁₈alkyl.

Some general examples of compounds described by the general structure(B) include a compound having the general structures (V)-(XXVIII):

In the case of 1,2,4-triazoles, there exist three tautomeric structures,as shown below:

Which tautomeric structure is prevailing depends on the substituents onthe triazole moiety and on the reaction conditions. As known to thosehaving ordinary skill in the art, typically, 1H-1,2,4-triazole is themost common tautomeric form, especially if an amino substituent isattached to the ring. Even though all three tautomeric structures can bepresent, all the generic structures and all the examples having1,2,4-triazole moiety are shown herein in one tautomeric form, such as4H-1,2,4-triazole, for simplicity and for the comparison with its directanalogues, such as examples containing 1,3,4-oxadiazole moiety. Usingonly 4H-tautomeric form to draw the structures for the sake ofsimplicity, does not imply that the compounds of the examples (30)-(74)shown below exist in that particular tautomeric form.

Some examples of particular compounds described by the general structure(B) include compounds having formulae (34)-(83):

Appropriately substituted 1,2,4-triazoles of the type (V) describedabove and illustrated by the general structure (B) can be synthesizedusing one of several reaction schemes, for example as shown in SchemesV, VI and VII below. The appropriate method can be chosen based on therequired substitution, availability of the starting materials and theease of synthesis.

1,2,4-triazoles of the type (VI) described above, can be made as shownin Scheme VIII or by an alternative route as outlined in Scheme IX.

Appropriately substituted 2-amino-1,3,4-oxadoazoles of type (VIII), canbe synthesized using one of several reaction schemes, for example, asshown by Schemes X, XI and XII.

Appropriately substituted 2-amino-1,3,4-oxadiazoles of type (IX), can besynthesized using one of several reaction schemes, for example, as shownby Schemes XIII and XIV.

Appropriately substituted 2-amino-1,3,4-thiadiazoles of type (XI), canbe prepared by a method which is outlined in Scheme XV.

Appropriately substituted 2-amino-1,3,4-thiadiazoles of type (XII), canbe prepared by a method which is outlined in Scheme XVI.

Appropriately substituted 3-amino-1,2,4-triazines of type (XXIII)-(XXV),can be prepared by a method which is outlined in Scheme XVII.

Appropriately substituted 3-amino-1,2,4-triazines of type(XXVI)-XXVIII), be prepared by a method which is outlined in SchemeXVIII.

The compounds and methods of the present invention, either whenadministered alone or in combination with other agents (e.g.,chemotherapeutic agents or protein therapeutic agents described below)are useful in treating a variety of disorders, including, but notlimited to, for example, cancer, eye disease, inflammation, psoriasis,and a viral infection. The kinds of cancer that can be treated include,but are not limited to, an alimentary/gastrointestinal tract cancer,colon cancer, liver cancer, skin cancer, breast cancer, ovarian cancer,prostate cancer, lymphoma, leukemia, kidney cancer, lung cancer, musclecancer, bone cancer, bladder cancer or brain cancer.

Embodiments of the present invention also provide articles ofmanufacture that can include a packaging material and a pharmaceuticalcomposition contained within the packaging material. The packagingmaterial can comprise a label which indicates that the pharmaceuticalcomposition can be used for treatment of one or more disordersidentified above.

The pharmaceutical composition can include a compound according to thepresent invention. In addition to a compound of the present invention,the pharmaceutical may also contain other therapeutic agents, and may beformulated, for example, by employing conventional solid or liquidvehicles or diluents, as well as pharmaceutical additives of a typeappropriate to the mode of desired administration (for example,excipients, binders, preservatives, stabilizers, flavors, etc.)according to techniques known in the art of pharmaceutical formulation.

Thus, in one embodiment, the invention provides a pharmaceuticalcomposition including a therapeutic agent and a compound of theinvention. The compound is present in a concentration effective to treatcancer.

The compounds of the invention may be formulated into therapeuticcompositions as natural or salt forms. Pharmaceutically acceptable,non-toxic salts include the base addition salts (formed with freecarboxyl or other anionic groups) which may be derived from inorganicbases such as, for example, sodium, potassium, ammonium, calcium, orferric hydroxides, and such organic bases as isopropylamine,trimethylamine, 2-ethylamino-ethanol, histidine, procaine, and the like.Such salts may also be formed as acid addition salts with any freecationic groups and will generally be formed with inorganic acids suchas, for example, hydrochloric, sulfuric, or phosphoric acids, or organicacids such as acetic, citric, p-toluenesulfonic, methanesulfonic acid,oxalic, tartaric, mandelic, and the like.

Salts of the invention can include amine salts formed by the protonationof an amino group with inorganic acids such as hydrochloric acid,hydrobromic acid, hydroiodic acid, sulfuric acid, phosphoric acid, andthe like. Salts of the invention can also include amine salts formed bythe protonation of an amino group with suitable organic acids, such asp-toluenesulfonic acid, acetic acid, methanesulfonic acid and the like.Additional excipients which are contemplated for use in the practice ofthe present invention are those available to those of ordinary skill inthe art, for example, those found in the United States Pharmacopeia Vol.XXII and National Formulary Vol. XVII, U.S. Pharmacopeia Convention,Inc., Rockville, Md. (1989), the relevant contents of which isincorporated herein by reference. In addition, polymorphs of theinvention compounds are included in the present invention.

Pharmaceutical compositions of the invention may be administered by anysuitable means, for example, orally, such as in the form of tablets,capsules, granules or powders; sublingually; buccally; parenterally,such as by subcutaneous, intravenous, intramuscular, intrathecal, orintracisternal injection or infusion techniques (e.g., as sterileinjectable aqueous or non-aqueous solutions or suspensions); nasallysuch as by inhalation spray; topically, such as in the form of a creamor ointment; or rectally such as in the form of suppositories; in dosageunit formulations containing non-toxic, pharmaceutically acceptablevehicles or diluents. The present compounds may, for example, beadministered in a form suitable for immediate release or extendedrelease. Immediate release or extended release may be achieved by theuse of suitable pharmaceutical compositions comprising the presentcompounds, or, particularly in the case of extended release, by the useof devices such as subcutaneous implants or osmotic pumps. The presentcompounds may also be administered liposomally.

In addition to primates, such as humans, a variety of other mammals canbe treated according to the method of the present invention. Forinstance, mammals including, but not limited to, cows, sheep, goats,horses, dogs, cats, guinea pigs, rats or other bovine, ovine, equine,canine, feline, rodent or murine species can be treated. However, themethod can also be practiced in other species, such as avian species(e.g., chickens).

The pharmaceutical compositions for the administration of the compoundsof this embodiment, either alone or in combination with othertherapeutic agents, may conveniently be presented in dosage unit formand may be prepared by any of the methods well known in the art ofpharmacy. All methods include bringing the active ingredient intoassociation with the carrier which constitutes one or more accessoryingredients. In general, the pharmaceutical compositions are prepared byuniformly and intimately bringing the active ingredient into associationwith a liquid carrier or a finely divided solid carrier or both, andthen, if necessary, shaping the product into the desired formulation. Inthe pharmaceutical composition the active object compound is included inan amount sufficient to produce the desired effect upon the process orcondition of diseases. The pharmaceutical compositions containing theactive ingredient may be in a form suitable for oral use, for example,as tablets, troches, lozenges, aqueous or oily suspensions, dispersiblepowders or granules, emulsions, hard or soft capsules, or syrups orelixirs.

Compositions intended for oral use may be prepared according to anymethod known to the art for the manufacture of pharmaceuticalcompositions and such compositions may contain one or more agentsselected from the group consisting of sweetening agents, flavoringagents, coloring agents and preserving agents in order to providepharmaceutically elegant and palatable preparations. Tablets contain theactive ingredient in admixture with non-toxic pharmaceuticallyacceptable excipients which are suitable for the manufacture of tablets.These excipients may be for example, inert diluents, such as calciumcarbonate, sodium carbonate, lactose, calcium phosphate or sodiumphosphate; granulating and disintegrating agents, for example, cornstarch, or alginic acid; binding agents, for example starch, gelatin oracacia, and lubricating agents, for example magnesium stearate, stearicacid or talc. The tablets may be uncoated or they may be coated by knowntechniques to delay disintegration and absorption in thegastrointestinal tract and thereby provide a sustained action over alonger period. For example, a time delay material such as glycerylmonostearate or glyceryl distearate may be employed. They may also becoated to form osmotic therapeutic tablets for control release.

Formulations for oral use may also be presented as hard gelatin capsuleswherein the active ingredient is mixed with an inert solid diluent, forexample, calcium carbonate, calcium phosphate or kaolin, or as softgelatin capsules wherein the active ingredient is mixed with water or anoil medium, for example peanut oil, liquid paraffin, or olive oil.

Aqueous suspensions contain the active materials in admixture withexcipients suitable for the manufacture of aqueous suspensions. Suchexcipients are suspending agents, for example sodiumcarboxymethylcellulose, methylcellulose, hydroxy-propylmethylcellulose,sodium alginate, polyvinyl-pyrrolidone, gum tragacanth and gum acacia;dispersing or wetting agents may be a naturally-occurring phosphatide,for example lecithin, or condensation products of an alkylene oxide withfatty acids, for example polyoxyethylene stearate, or condensationproducts of ethylene oxide with long chain aliphatic alcohols, forexample heptadecaethyleneoxycetanol, or condensation products ofethylene oxide with partial esters derived from fatty acids and ahexitol such as polyoxyethylene sorbitol monooleate, or condensationproducts of ethylene oxide with partial esters derived from fatty acidsand hexitol anhydrides, for example polyethylene sorbitan monooleate.Also useful as a solubilizer is polyethylene glycol, for example. Theaqueous suspensions may also contain one or more preservatives, forexample ethyl, or n-propyl, p-hydroxybenzoate, one or more coloringagents, one or more flavoring agents, and one or more sweetening agents,such as sucrose or saccharin.

Oily suspensions may be formulated by suspending the active ingredient avegetable oil, for example arachis oil, olive oil, sesame oil or coconutoil, or in a mineral oil such as liquid paraffin. The oily suspensionsmay contain a thickening agent, for example beeswax, hard paraffin orcetyl alcohol. Sweetening agents such as those set forth above, andflavoring agents may be added to provide a palatable oral preparation.These compositions may be preserved by the addition of an anti-oxidantsuch as ascorbic acid.

Dispersible powders and granules suitable for preparation of an aqueoussuspension by the addition of water provide the active ingredient inadmixture with a dispersing or wetting agent, suspending agent and oneor more preservatives. Suitable dispersing or wetting agents andsuspending agents are exemplified by those already mentioned above.Additional excipients, for example sweetening, flavoring and coloringagents, may also be present.

Syrups and elixirs may be formulated with sweetening agents, for exampleglycerol, propylene glycol, sorbitol or sucrose. Such formulations mayalso contain a demulcent, a preservative and flavoring and coloringagents.

The pharmaceutical compositions may be in the form of a sterileinjectable aqueous or oleagenous suspension. This suspension may beformulated according to the known art using those suitable dispersing orwetting agents and suspending agents which have been mentioned above.The sterile injectable preparation may also be a sterile injectablesolution or suspension in a parenterally-acceptable diluent or solventor cosolvent or complexing agent or dispersing agent or excipient orcombination thereof, for example 1,3-butanediol, polyethylene glycols,polypropylene glycols, ethanol or other alcohols, povidones, variousbrands of TWEEN surfactant, sodium dodecyl sulfate, sodium deoxycholate,dimethylacetamide, polysorbates, poloxamers, cyclodextrins, lipids, andexcipients such as inorganic salts (e.g., sodium chloride), bufferingagents (e.g., sodium citrate, sodium phosphate), and sugars (e.g.,saccharose and dextrose). Among the acceptable vehicles and solventsthat may be employed are water, dextrose solutions, Ringer's solutionsand isotonic sodium chloride solution. In addition, sterile, fixed oilsare conventionally employed as a solvent or suspending medium. For thispurpose any bland fixed oil may be employed including synthetic mono- ordiglycerides. In addition, fatty acids such as oleic acid find use inthe preparation of injectables.

Depending on the condition being treated, these pharmaceuticalcompositions may be formulated and administered systemically or locally.Techniques for formulation and administration may be found in the latestedition of “Remington's Pharmaceutical Sciences” (Mack Publishing Co,Easton, Pa.). Suitable routes may, for example, include oral ortransmucosal administration; as well as parenteral delivery, includingintramuscular, subcutaneous, intramedullary, intrathecal,intraventricular, intravenous, intraperitoneal, or intranasaladministration. For injection, the pharmaceutical compositions of theinvention may be formulated in aqueous solutions, preferably inphysiologically compatible buffers such as Hanks' solution, Ringer'ssolution, or physiologically buffered saline. For tissue or cellularadministration, penetrants appropriate to the particular harrier to bepermeated are used in the formulation. Such penetrants are generallyknown in the art. Pharmaceutical formulations for parenteraladministration include aqueous solutions of the active compounds inwater-soluble form. Additionally, suspensions of the active compoundsmay be prepared as appropriate oily injection suspensions. Suitablelipophilic solvents or vehicles include fatty oils such as sesame oil,or synthetic fatty acid esters, such as ethyl oleate or triglycerides,or liposomes. Aqueous injection suspensions may contain substances thatincrease the viscosity of the suspension, such as sodium carboxymethylcellulose, sorbitol, or dextran. Optionally, the suspension may alsocontain suitable stabilizers or agents that increase the solubility ofthe compounds to allow for the preparation of highly concentratedsolutions.

The compounds of the present invention may also be administered in theform of suppositories for rectal administration of the drug. Thesecompositions can be prepared by mixing the drug with a suitablenon-irritating excipient which is solid at ordinary temperatures butliquid at the rectal temperature and will therefore melt in the rectumto release the drug. Such materials are cocoa butter and polyethyleneglycols.

For topical use, creams, ointments, jellies, solutions or suspensions,etc., containing the compounds of the present invention are employed.(For purposes of this application, topical application shall includemouthwashes and gargles).

In one embodiment, the invention compounds are administered incombination with an anti-inflammatory agent, antihistamines,chemotherapeutic agent, immunomodulator, therapeutic antibody or aprotein kinase inhibitor, e.g., a tyrosine kinase inhibitor, to asubject in need of such treatment. While not wanting to be limiting,chemotherapeutic agents include antimetabolites, such as methotrexate,DNA cross-linking agents, such as cisplatin/carboplatin; alkylatingagents, such as canbusil; topoisomerase I inhibitors such asdactinomycin; microtubule inhibitors such as taxol (paclitaxol), and thelike. Other chemotherapeutic agents include, for example, a vincaalkaloid, mitomycin-type antibiotic, bleomycin-type antibiotic,antifolate, colchicine, demecolcine, etoposide, taxane, anthracyclineantibiotic, doxorubicin, daunorubicin, caminomycin, epirubicin,idarubicin, mitoxanthrone, 4-dimethoxy-daunomycin, 11-deoxydaunorubicin,13-deoxydaunorubicin, adriamycin-14-benzoate, adriamycin-14-octanoate,adriamycin-14-naphthaleneacetate, amsacrine, carmustine,cyclophosphamide, cytarabine, etoposide, lovastatin, melphalan,topetecan, oxalaplatin, chlorambucil, methotrexate, lomustine,thioguanine, asparaginase, vinblastine, vindesine, tamoxifen, ormechlorethamine. While not wanting to be limiting, therapeuticantibodies include antibodies directed against the HER2 protein, such astrastuzumab; antibodies directed against growth factors or growth factorreceptors, such as bevacizumab, which targets vascular endothelialgrowth factor, and OSI-774, which targets epidermal growth factor;antibodies targeting integrin receptors, such as Vitaxin (also known asMEDI-522), and the like. Classes of anticancer agents suitable for usein compositions and methods of the present invention include, but arenot limited to: 1) alkaloids, including, microtubule inhibitors (e.g.,Vincristine, Vinblastine, and Vindesine, etc.), microtubule stabilizers(e.g., Paclitaxel [Taxol], and Docetaxel, Taxotere, etc.), and chromatinfunction inhibitors, including, topoisomerase inhibitors, such as,epipodophyllotoxins (e.g., Etoposide [VP-16], and Teniposide [VM-26],etc.), and agents that target topoisomerase I (e.g., Camptothecin andIsirinotecan [CPT-11], etc.); 2) covalent DNA-binding agents [alkylatingagents], including, nitrogen mustards (e.g., Mechlorethamine,Chlorambucil, Cyclophosphamide, Ifosphamide, and Busulfan [Myleran],etc.), nitrosoureas (e.g., Carmustine, Lomustine, and Semustine, etc.),and other alkylating agents (e.g., Dacarbazine, Hydroxymethylmelamine,Thiotepa, and Mitocycin, etc.); 3) noncovalent DNA-binding agents[antitumor antibiotics], including, nucleic acid inhibitors (e.g.,Dactinomycin [Actinomycin D], etc.), anthracyclines (e.g., Daunorubicin[Daunomycin, and Cerubidine], Doxorubicin [Adriamycin], and Idarubicin[Idamycin], etc.), anthracenediones (e.g., anthracycline analogues, suchas, [Mitoxantrone], etc.), bleomycins (Blenoxane), etc., and plicamycin(Mithramycin), etc.; 4) antimetabolites, including, antifolates (e.g.,Methotrexate, Folex, and Mexate, etc.), purine antimetabolites (e.g.,6-Mercaptopurine [6-MP, Purinethol], 6-Thioguanine [6-TG], Azathioprine,Acyclovir, Ganciclovir, Chlorodeoxyadenosine, 2-Chlorodeoxyadenosine[CdA], and 2′-Deoxycoformycin [Pentostatin], etc.), pyrimidineantagonists (e.g., fluoropyrimidines [e.g., 5-fluorouracil (Adrucil),5-fluorodeoxyuridine (FdUrd) (Floxuridine)] etc.), and cytosinearabinosides (e.g., Cytosar [ara-C] and Fludarabine, etc.); 5) enzymes,including, L-asparaginase; 6) hormones, including, glucocorticoids, suchas, antiestrogens (e.g., Tamoxifen, etc.), nonsteroidal antiandrogens(e.g., Flutamide, etc.), and aromatase inhibitors (e.g., anastrozole[Arimidex], etc.); 7) platinum compounds (e.g., Cisplatin andCarboplatin, etc.); 8) monoclonal antibodies conjugated with anticancerdrugs, toxins, and/or radionuclides, etc.; 9) biological responsemodifiers (e.g., interferons [e.g., IFN-.alpha., etc.] and interleukins[e.g., IL-2, etc.], etc.); 10) adoptive immunotherapy; 11) hematopoieticgrowth factors; 12) agents that induce tumor cell differentiation (e.g.,all-trans-retinoic acid, etc.); 13) gene therapy techniques; 14)antisense therapy techniques; 15) tumor vaccines; 16) therapies directedagainst tumor metastases (e.g., Batimistat, etc.); and 17) inhibitors ofangiogenesis.

The pharmaceutical composition and method of the present invention mayfurther comprise other therapeutically active compounds as noted hereinwhich are usually applied in the treatment of the above mentionedpathological conditions. Examples of other therapeutic agents includethe following: cyclosporins (e.g., cyclosporin A), CTLA4-Ig, antibodiessuch as ICAM-3, anti-IL-2 receptor (Anti-Tac), anti-CD45RB, anti-CD2,anti-CD3 (OKT-3), anti-CD4, anti-CD 80, anti-CD86, agents blocking theinteraction between CD40 and gp39, such as antibodies specific for CD40and/or gp39 (i.e., CD154), fusion proteins constructed from CD40 andgp39 (CD40Ig and CD8gp39), inhibitors, such as nuclear translocationinhibitors, of NF-kappa B function, such as deoxyspergualin (DSG),cholesterol biosynthesis inhibitors such as HMG CoA reductase inhibitors(lovastatin and simvastatin), non-steroidal antiinflammatory drugs(NSAIDs) such as ibuprofen and cyclooxygenase inhibitors such asrofecoxib, steroids such as prednisone or dexamethasone, gold compounds,antiproliferative agents such as methotrexate, FK506 (tacrolimus,Prograf), mycophenolate mofetil, cytotoxic drugs such as azathioprineand cyclophosphamide, TNF-a inhibitors such as tenidap, anti-TNFantibodies or soluble TNF receptor, and rapamycin (sirolimus orRapamune) or derivatives thereof.

Other agents that may be administered in combination with inventioncompounds include protein therapeutic agents such as cytokines,immunomodulatory agents and antibodies. As used herein the term“cytokine” encompasses chemokines, interleukins, lymphokines, monokines,colony stimulating factors, and receptor associated proteins, andfunctional fragments thereof. As used herein, the term “functionalfragment” refers to a polypeptide or peptide which possesses biologicalfunction or activity that is identified through a defined functionalassay.

The cytokines include endothelial monocyte activating polypeptide II(EMAP-II), granulocyte-macrophage-CSF (GM-CSF), granulocyte-CSF (G-CSF),macrophage-CSF (M-CSF), IL-1, IL-2, IL-3, IL-4, IL-5, IL-6, IL-12, andIL-13, interferons, and the like and which is associated with aparticular biologic, morphologic, or phenotypic alteration in a cell orcell mechanism.

When other therapeutic agents are employed in combination with thecompounds of the present invention they may be used for example inamounts as noted in the Physician Desk Reference (PDR) or as otherwisedetermined by one having ordinary skill in the art.

In the treatment or prevention of conditions which involve cellularproliferation, an appropriate dosage level can generally be betweenabout 0.01 and about 1000 mg per 1 kg of patient body weight per daywhich can be administered in single or multiple doses. For example, thedosage level can be between about 0.01 and about 250 mg/kg per day; morenarrowly, between about 0.5 and about 100 mg/kg per day. A suitabledosage level can be between about 0.01 and about 250 mg/kg per day,between about 0.05 and about 100 mg/kg per day, or between about 0.1 andabout 50 mg/kg per day, or about 1.0 mg/kg per day. For example, withinthis range the dosage can be between about 0.05 and about 0.5 mg/kg perday, or between about 0.5 and about 5 mg/kg per day, or between about 5and about 50 mg/kg per day. For oral administration, the compositionscan be provided in the form of tablets containing between about 1.0 andabout 1,000 mg of the active ingredient, for example, about 1.0, about5.0, about 10.0, about 15.0, about 20.0, about 25.0, about 50.0, about75.0, about 100.0, about 150.0, about 200.0, about 250.0, about 300.0,about 400.0, about 500.0, about 600.0, about 750.0, about 800.0, about900.0, and about 1,000.0 mg of the active ingredient for the symptomaticadjustment of the dosage to the patient to be treated. The compounds canbe administered on a regimen of 1 to 4 times per day, such as once ortwice per day. There may be a period of no administration followed byanother regimen of administration.

It will be understood, however, that the specific dose level andfrequency of dosage for any particular patient may be varied and willdepend upon a variety of factors including the activity of the specificcompound employed, the metabolic stability and length of action of thatcompound, the age, body weight, general health, sex, diet, mode and timeof administration, rate of excretion, drug combination, the severity ofthe particular condition, and the host undergoing therapy.

Compounds of the present invention can be used, alone or in combinationwith an effective amount of a therapeutic antibody or chemicallyattached to a tumor tissue targeting antibody (or therapeutic fragmentthereof), a chemotherapeutic or an immunotoxic agent, for treatment oftumors. Illustrative examples of chemotherapeutic agents that can beused for this purpose include doxorubicin, docetaxel, or taxol. Itshould be further understood that the invention includes combinationtherapy including a compound of the invention, including but not limitedto vasculostatic agents, such as tyrosine, serine or threonine kinaseinhibitors, for example, Src-family inhibitors, and any chemotherapeuticagent or therapeutic antibody.

The present invention also provides screening assays using appropriatecells which express any kinases within the MAPK pathway. Such cellsinclude cells from mammals, yeast, Drosophila or E. coli. For example,cells which express the Raf polypeptide or any kinase downstream of Rafsuch as MEK or ERK1/2 or respond to Raf polypeptide or MAPK pathwaypolypeptides are contacted with a test compound to observe binding, orstimulation or inhibition of a functional response. The cells which arecontacted with the candidate compound are compared with the same cellswhich are not contacted for Raf polypeptide or MAPK pathway polypeptideactivity

This invention contemplates the treatment and/or amelioration of suchdiseases by administering a MAPK pathway polypeptides inhibiting amountof a compound. Without wishing to be bound by any particular theory ofthe functioning of the MAPK pathway polypeptides, it is believed thatamong the useful inhibitors of MAPK pathway polypeptides function arethose compounds which inhibit the kinase activity of the MAPK pathwaypolypeptides. Other sites of inhibition are, of course, possible owingto its position in a signal transduction cascade. Inhibitors ofprotein-protein interactions between, for example Raf polypeptide orMAPK pathway polypeptides and other factors could lead to thedevelopment of pharmaceutical agents for the modulation of Rafpolypeptide or MAPK pathway polypeptides activity.

Targeting an allosteric site of the protein is a very promising approachfor pharmaceutical intervention. Further, the traditional approach ofinhibiting various protein kinases includes targeting the ATP bindingsite. The invention is not meant to be limited by any particularmechanism of inhibition. The assays of the invention may test binding ofa candidate compound wherein adherence to the cells bearing the Rafpolypeptide or MAPK pathway polypeptides is detected by means of a labeldirectly or indirectly associated with the candidate compound or in anassay involving competition with a labeled competitor. Further, theseassays may test whether the candidate compound results in a signalgenerated by activation of MAPK pathway polypeptides, using detectionsystems appropriate to the cells bearing the Raf poly-peptide or MAPKpathway polypeptides. Inhibitors of activation are generally assayed inthe presence of a known agonist and the effect on activation by theagonist by the presence of the candidate compound is observed. Standardmethods for conducting such screening assays are well understood in theart and are also illustrated in the examples below (e.g., direct andraf1-MEK1 assays or MAPK pathway cellular assays).

The following examples are provided to further illustrate the advantagesand features of the present invention, but are not intended to limit thescope of the invention.

Example 1 General Methods

Example 1 describes general synthetic procedures that were used to makethe compounds described in the subsequent examples. All solvents wereused without further purification. Reactions can be usually conductedwithout an inert gas atmosphere unless specified otherwise. The reportedyields are based on unoptimized conditions and single test runs. Theyields can be optimized by changing the reaction conditions, such assolvent, use of base or acid, temperature, use of catalyst and the timeof the reaction. Microwave reactions were run in Emrys™ Process vials(2-5 mL) using Initiator module (Biotage/Personal chemistry). All ¹H NMRwere run on a 500 MHz Bruker NMR or Bruker Avance 400 MHz NMR. Chemicalshifts are reported in delta (δ) units, parts per million (ppm)downfield from tetramethylsilane. Coupling constants are reported inhertz (Hz). A Waters LC/MS system is used in identity and purityanalysis. This system includes a 2795 separation module, a 996photodiode array detector and a ZQ2000 mass spectrometer, A Zorbax SBcolumn (150×4.6 mm 3.5μ, Agilent Technologies) was used for the LC.Column temperature was 40° C. Compounds were separated using gradientelution with mobile phases of water (0.05% TFA (A)) and acetonitrile(0.05% TFA (B)). Flow rate was 1 mL/min. The gradient program used inseparation was 0-15 min: 5-60% B; 15-15.5 min: 60-100% B; 15.5-17 min:100% B.

Example 2 Synthesis of 3-amino-benzo[1,2,4]triazine-7-ol-1-oxide

7.7 g (0.05 mol) of 4-amino-3-nitrophenol was dissolved in 20 mL ofglacial acetic acid and the resulting bright-red solution was heated toapproximately 100° C. in 500 mL round-bottom flask equipped with a longcondenser. To this solution was added a solution of 16.81 g (8.0equivalent, 0.4 mol) of cyanamide in 20 mL of concentrated hydrochloricacid. In approximately 5-10 min the reaction mixture started to boilvigorously, so the heating was removed and it was stirred withoutheating until boiling subsided. Then the heating was reapplied and thereaction mixture was refluxed for 48 hrs. Then 150 mL of 30% NaOH wasadded and the resulting dark-reddish solution was refluxed foradditional 3 hrs. Then it was cooled down to room temperature anddark-red slurry was formed. The red precipitate was filtered,re-dissolved in 200 mL of water and 1N HCl was added in portions withstirring until pH reached 5-4. The solution changed color from dark-redinto light-yellow and a light-yellow fine precipitate was formed. Theprecipitate was filtered, washed twice with 50 mL of water, twice with50 mL of acetonitrile and finally twice with 50 mL of diethyl ether anddried in vacuum to give 4.6 g of a bright-yellow solid. Yield: 51.7%.

¹H NMR (DMSO-d₆): δ 6.96 (s, 2H), 7.35-7.38 (m, 2H), 7.45-7.47 (m, 1H),10.36 (s, 1H).

Example 3 Synthesis of 3-amino-benzo[1,2,4]triazine-7-ol

4.6 g (25.82 mmol) of 3-amino-benzo[1,2,4]triazine-7-ol-1-oxide wasdissolved in 200 mL of 1:1 mixture of dimethylformamide and methanol.0.5 g of 10% Pd/C was added to this solution and H₂ gas was bubbledthrough the solution for 3 hours. The progress of the reaction wasmonitored by TLC, using a 9:1 mixture of dichloromethane/methanol as aneluent and a UV lamp. The starting material is highly fluorescent underUV, while the product is not. When the reaction was complete, theresulting dark solution was filtered through a short pad of silica geland solvent was removed in vacuum to produce a dirty-brown solid. 40 mLof ethyl acetate and 40 mL of methanol were added to the solid and theresulting suspension was heated to reflux for about 10 min. Then thesuspension was allowed to cool down to ambient temperature. The solidwas collected by filtration, washed with 40 mL of ethyl acetate, 40 mLof diethyl ether and dried in vacuum to yield 3.2 g of the product in aform of a greenish solid. Yield: 76%.

¹H NMR (DMSO-d₆): δ 7.18 (s, 1H), 7.36 (d, J=2.6 Hz, 1H), 7.40-7.42 (dd,J₁=9.1 Hz, J₂=2.6 Hz, 1H), 7.45-7.46 (d, J=9.1 Hz, 1H).

Example 4 Synthesis of 4-chloro-pyridine-2-carboxylic acid methyl esterhydrochloride salt

2.4 mL (0.031 mol, 0.16 equivalent) of anhydrous N,N-dimethylformamidewas added dropwise to 72 mL (1.23 mol, 5.0 equivalent) of thionylchloride in a temperature range of 40-50° C. under argon blanket. Thesolution was stirred at this temperature for 10 min, then 24.0 g (0.195mol, 1.0 equivalent) of picolinic acid was added slowly in portions. Thereaction mixture was heated at 70-75° C. with a reflux condenser underargon for 30 hours. Evolution of SO₂ gas was observed. The reactionmixture changed colors from green to orange, then to purple over 2hours, then resulted in an orange solution with a yellow precipitate. Itwas cooled down to ambient temperature and 150 mL of anhydrous toluenewas added. The suspension was concentrated to about 50 mL total onrotovap. This process was repeated three times.

The resulting orange suspension was cooled down to −20° C. and 200 mL ofmethanol was added. The reaction mixture was left to stir at ambienttemperature for 18 hours. Then the clear-yellow solution was transferredinto a round-bottom flask and solvent was removed in vacuum. Theresulting yellow solid was dissolved with heating to 50° C. in 50 mL ofmethanol, upon cooling 300 mL of diethyl ether were added. The solutionwas left to stand at 0° C. for 18 hours. The white precipitate thatformed was collected by filtration, washed extensively with diethylether and dried in vacuum to yield 29.05 g of the product as a whitefluffy solid. Yield: 71.5%.

¹H NMR (DMSO-d₆): δ 3.88 (s, 3H), 7.81-7.83 (dd, J₁=1.9 Hz, J₂=5.4 Hz,1H), 8.06-8.07 (d, J=1.9 Hz, 1H), 8.68-8.69 (d, J=5.4 Hz, 1H).

Example 5 Synthesis of 4-chloro-pyridine-2-carboxylic acid methyl amide

A suspension of 17.8 g (0.103 mol, 1 eq) of4-chloro-pyridine-2-carboxylic acid methyl ester hydrochloride in 15 mL,of methanol was cooled to 0° C. and slowly treated with a 2.0 M solutionof methylamine in tetrahydrofuran at a rate that kept internaltemperature below 5° C. The reaction mixture was stirred at 0° C. for 2hours, then slowly allowed to warm up to ambient temperature and stirredfor 18 hours. Solvent was removed in vacuum, approx. 200 mL of ethylacetate was added and the resulting suspension was filtered. Theprecipitate was washed with 100 mL of ethyl acetate. The combined ethylacetate solutions were washed three times with 100 mL of brine and driedover sodium sulfate. Solvent was removed in vacuum to yield 14.16 g ofthe product as orange oil. Yield: 80.5%.

¹H NMR (DMSO-d₆): δ 2.81-2.82 (d, J=4.8 Hz, 3H), 7.73-7.75 (dd, J₁=2.1Hz, J₂=5.4 Hz, 1H), 8.00-8.01 (d, J=2.1 Hz, 1H), 8.60-8.61 (d, J=5.4 Hz,1H), 8.84 (q, J=4.8 Hz, 1H).

Example 6 Synthesis of4-(3-amino-benzo[1,2,4]triazin-7-yloxy)-pyridine-2-carboxylic acidmethylamide

3.2 g (19.73 mmol) of 3-amino-benzo[1,2,4]triazine-7-ol was dissolved in80 mL of anhydrous dimethylformamide under argon atmosphere. 2.44 g(21.71 mmol, 1.1 equivalent) of solid potassium tert-butoxide was addedto the solution. The resulting dark-red mixture was heated to about 100°C. and stirred at that temperature for 15 min. A solution of 3.7 g(21.71 mmol, 1.1 equivalent) of 4-chloro-pyridine-2-carboxylic acidmethylamide in 10 mL of anhydrous dimethylformamide was added, followedby 3.28 g (23.68 mmol, 1.2 equivalent) of anhydrous K₂CO₃. The reactionmixture was heated at 140° C. for 30 hrs. The progress of the reactionwas monitored by LC/MS. Then it was allowed to cool down to ambienttemperature. The resulting dark-brown slurry was poured into 500 mL ofwater and 100 mL of ethyl acetate. The formed precipitate was collectedby filtration, washed with 50 mL of water, 50 mL of methanol, 50 mL ofdiethyl ether and dried in vacuum to produce 3.22 g of the product as adirty-yellow solid. The filtrate was extracted four times with 100 mL ofethyl acetate. The combined extracts were washed 3 times with 100 mL ofwater, then with brine and dried over anhydrous sodium sulfate. Solventwas removed in vacuum to yield additional 1.2 g of the product in a formof a yellow solid. Yield: 75% combined.

¹H NMR (DMSO-d₆): δ 2.78-2.79 (d, J=4.8 Hz, 3H), 7.25-7.27 (dd, J₁=2.6Hz, J₂=5.6 Hz, 1H), 7.50-7.51 (d, J=2.6 Hz, 1H), 7.66-7.68 (d, J=9.2 Hz,1H), 7.71-7.73 (dd, J₁=2.7 Hz, J₂=9.2 Hz, 1H), 7.71 (s, 2H), 8.03-8.05(d, J=2.7 Hz, 1H), 8.54-8.55 (d, J=5.6 Hz, 1H), 8.78-8.80 (q, J=4.8 Hz,1H).

Example 7 Synthesis of4-[3-(4-chloro-3-trifluoromethyl-phenylamino)-benzo[1,2,4]triazin-7-yloxy]-pyridine-2-carboxylicacid methylamide trifluoroacetate salt

To a vial with 2 mL of anhydrous dimethylformamide under argonatmosphere were added 15.4 mg (0.0168 mmol, 0.05 equivalent) oftris(dibenzyllideneatone) dipalladium(0), 21.0 mg (0.033 mmol, 0.1equivalent) of BINAP, 220.0 mg (0.675 mmol, 2.0 equivalent) of anhydrouscesium carbonate, 175.1 mg (0.675 mmol, 2.0 equivalent) of5-bromo-2-chlorobenzotrifluoride, and 100 mg (0.337 mmol, 1.0equivalent) of4-(3-amino-benzo[1,2,4]triazin-7-yloxy)-pyridine-carboxylic acidmethylamide, in that particular order. Argon gas was bubbled through themixture for 5 min. Then the vial was capped and the reaction mixture washeated to 120° C. with stirring under argon atmosphere for 18 hours. Atthis point LC/MS indicated about 40% conversion to the product. Aspointed out in the previous examples, the longer reaction times resultin the formation of the by-product and partial decomposition. So, thereaction mixture was allowed to cool down to ambient temperature,filtered through 0.3 μm syringe filter and purified by reverse-phaseprep-HPLC using acetonitrile/water mixture with 0.1% of TFA.

ESI-MS: [M+H]⁺, 475, 477. ¹H NMR (DMSO-d₆): δ 2.79-2.90 (d, J=4.8 Hz,3H), 7.31-7.33 (dd, J₁=2.4 Hz, J₂=5.4 Hz, 1H), 7.55-7.55 (d, J=2.4 Hz,1H), 7.73-7.75 (d, J=8.9 Hz, 1H), 7.89-7.91 (dd, J₁=2.5 Hz, J₂+9.2 Hz,1H), 7.94-7.95 (d, J=9.2 Hz, 1H), 8.24 (d, J=2.5 Hz, 2H), 8.25-8.27 (dd,J₁=2.6 Hz, J₂=8.9 Hz, 1H), 8.55 (d, J=2.6 Hz, 1H), 8.60 (d, J=5.4 Hz,1H), 8.81-8.82 (q, J=4.8 Hz, 1H), 11.36 (s, 1H).

Example 8 Synthesis of4-[3-(4-chloro-phenylamino)-benzo[1,2,4]triazin-7-yloxy]-pyridine-2-carboxylicacid methylamide trifluoroacetate salt

To a vial with 2 mL of anhydrous dimethylformamide under argonatmosphere were added 15.4 mg (0.0168 mmol, 0.05 equivalent) oftris(dibenzyllideneatone) dipalladium(0), 21.0 mg (0.033 mmol, 0.1equivalent) of BINAP, 220.0 mg (0.675 mmol, 2.0 equivalent) of anhydrouscesium carbonate, 161.0 mg (0.675 mmol, 2.0 equivalent) of1-chloro-4-iodobenzene, and 100 mg (0.337 mmol, 1.0 equivalent) of4-(3-amino-benzo[1,2,4]triazin-7-yloxy)-pyridine-carboxylic acidmethylamide, in that particular order. Argon gas was bubbled through themixture for 5 min. Then the vial was capped and the reaction mixture washeated to 120° C. with stirring under argon atmosphere for 18 hours. Atthis point LC/MS indicated about 30% conversion to the product. As itwas observed from previous examples, the longer reaction times result inthe formation of the by-product and partial decomposition. The reactionmixture was allowed to cool down to ambient temperature, filtered,through 0.22μ syringe filter and purified by reverse-phase preperativeHPLC using acetonitrile/water mixture with 0.1% of TFA.

ESI-MS: [M+H]⁺, 407, 409. ¹H NMR (DMSO-d₆): δ 2.79-2.80 (d, J=4.88 Hz,3H), 7.30-7.32 (dd, J₁=2.6 Hz, J₂=5.6 Hz, 1H), 7.44-7.46 (d, J=6.8 Hz,2H), 7.54-7.55 (d, J=2.6 Hz, 1H), 7.84-7.87 (dd, J₁=2.6 Hz, J₂=9.05 Hz,1H), 7.92-7.94 (d, J=9.05 Hz, 1H), 8.00-8.01 (d, J=6.8 Hz, 2H),8.19-8.20 (d, J=2.6 Hz, 1H), 8.58-8.59 (d, J=5.6 Hz, 1H), 8.81-8.82 (q,J=4.88 Hz, 1H), 11.08 (s, 1H).

Example 9 Synthesis of4-[3-(3-trifluoromethyl-benzenesulfonylamino)-benzo[1,2,4]triazin-7-yloxy]-pyridine-2-carboxylicacid methyl amide trifluoroacetate salt

100 mg (0.337 mmol, 1.0 equivalent) of4-(3-amino-benzo[1,2,4]triazin-7-yloxy)-pyridine-carboxylic acidmethylamide were dissolved in 2 mL of anhydrous dimethylformamide withheating to about 100° C. 45.4 mg (0.405 mmol, 1.2 equivalent) of solidtert-BuOK was added to the solution. The resulting dark-red solution wasstirred at 100° C. for 30 min, then it was allowed to cool down toambient temperature. 100 mg (0.405 mmol, 1.2 equivalent) of3-trifluoromethyl-benzenesulfonyl chloride was added to the mixture viaa syringe. It was allowed to stir at ambient temperature for 2 hours.The reaction mixture was filtered through 0.22μ syringe filter andpurified by reverse-phase preperative HPLC using acetonitrile/watermixture with 0.1% of TFA.

ESI-MS: [M+H]⁺, 505, 506, 507. ¹H NMR (DMSO-d₆): δ 2.78-2.79 (d, J=4.8Hz, 3H), 7.29-7.31 (dd, J₁=2.6 Hz, J₂=5.7 Hz, 1H), 7.56 (d, J=2.6 Hz,1H), 7.87-7.90 (t, J=7.8 Hz, 1H), 7.93-7.94 (d, J=9.2 Hz, 1H), 7.97-7.99(dd, J₁=2.6 Hz, J₂=9.2 Hz, 1H), 8.06-8.08 (d, J=7.8 Hz, 1H), 8.21 (d,J=2.5 Hz, 1H), 8.43-8.46 (m, 2H), 8.57-8.58 (d, J=5.7 Hz, 1H), 8.80-8.81(q, J=4.8 Hz, 1H).

Example 10 Synthesis of4-[3-(3-trifluoromethyl-benzoylamino)-benzo[1,2,4]triazin-7-yloxy]-pyridine-2-carboxylicacid methyl amide trifluoroacetate salt

100 mg (0.337 mmol, 1.0 equivalent) of4-(3-amino-benzo[1,2,4]triazin-7-yloxy)-pyridine-carboxylic acidmethylamide were dissolved in 2 mL of anhydrous dimethylformamide withheating to about 100° C. 45.4 mg (0.405 mmol, 1.2 equivalent) of solidtert-BuOK was added to the solution. The resulting dark-red solution wasstirred at 100° C. for 30 min, then it was allowed to cool down toambient temperature. 84.5 mg (0.405 mmol, 1.2 equivalent) of3-trifluoromethyl-benzoyl chloride was added to the mixture via asyringe. It was allowed to stir at ambient temperature for 2 hours. Thereaction mixture was filtered through 0.22μ syringe filter and purifiedby reverse-phase prep-HPLC using acetonitrile/water mixture with 0.1% ofTFA.

ESI-MS: [M+H]⁺, 469, 470. ¹H NMR (DMSO-d₆): δ 2.80-2.81 (d, J=4.88 Hz,3H), 7.37-7.39 (dd, J₁=2.6 Hz, J₂=5.7 Hz, 1H), 7.64 (d, J=2.6 Hz, 1H),7.81-7.84 (t, J=7.8 Hz, 1H), 8.03-8.06 (m, 2H), 8.15-8.17 (d, J=9.2 Hz,1H), 8.33-8.34 (d, J=2.6 Hz, 1H), 8.36-8.38 (d, J=8.0 Hz, 1H), 8.45 (s,1H), 8.62-8.63 (d, J=5.7 Hz, 1H), 8.84-8.85 (q, J=4.88 Hz, 1H), 12.29(s, 1H).

Example 11 Synthesis of4-[4-(trifluoromethoxy-benzoylamino)-benzo[1,2,4]triazin-7-yloxy]-pyridine-2-carboxylicacid methyl amide trifluoroacetate salt

100 mg (0.337 mmol, 1.0 equivalent) of4-(3-amino-benzo[1,2,4]triazin-7-yloxy)-pyridine-carboxylic acidmethylamide were dissolved in 2 mL of anhydrous DMF with heating toabout 100° C. 45.4 mg (0.405 mmol, 1.2 equivalent) of solid t-BuOK wasadded to the solution. The resulting dark-red solution was stirred at100° C. for 30 min, then it was allowed to cool down to ambienttemperature. 64 μL (91.0 mg, 0.405 mmol, 1.2 equivalent) of4-trifluoromethoxy-benzoyl chloride was added to the mixture via asyringe. It was allowed to stir at ambient temperature for 2 hours. Thereaction mixture was filtered through 0.22μ syringe filter and purifiedby reverse-phase prep-HPLC using acetonitrile/water mixture with 0.1% ofTFA as a solvent system.

ESI-MS: [M+H]⁺, 485, 486. ¹H NMR (DMSO-d₆): δ 2.80-2.81 (d, J=4.8 Hz,3H), 7.37-7.38 (dd, J₁=2.6 Hz, J₂=5.6 Hz, 1H), 7.55-7.57 (d, J=8.8 Hz,2H), 7.63-7.64 (d, J=2.6 Hz, 1H), 8.03-8.05 (dd, J₁=2.7 Hz, J₂=9.1 Hz,1H), 8.14-8.15 (d, J=9.1 Hz, 1H), 8.20-8.22 (d, J=8.8 Hz, 2H), 8.32-8.33(d, J=2.7 Hz, 1H), 8.83-8.84 (q, J=4.8 Hz, 1H), 12.12 (s, 1H).

Example 12 Synthesis of4-[3-(trifluoromethoxy-benzoylamino)-benzo[1,2,4]triazin-7-yloxy]-pyridine-2-carboxylicacid methyl amide trifluoroacetate salt

100 mg (0.337 mmol, 1.0 equivalent) of4-(3-amino-benzo[1,2,4]triazin-7-yloxy)-pyridine-carboxylic acidmethylamide were dissolved in 2 mL of anhydrous DMF with heating toabout 100° C. 45.4 mg (0.405 mmol, 1.2 equivalent) of solid t-BuOK wasadded to the solution. The resulting dark-red solution was stirred at100° C. for 30 min, then it was allowed to cool down to ambienttemperature. 64 μL (91.0 mg, 0.405 mmol, 1.2 equivalent) of3-trifluoromethoxy-benzoyl chloride was added to the mixture via asyringe. It was allowed to stir at ambient temperature for 2 hours. Thereaction mixture was filtered through 0.22μ syringe filter and purifiedby reverse-phase prep-HPLC using acetonitrile/water mixture with 0.1% ofTFA as a solvent system.

ESI-MS: [M+H]⁺, 485, 486. ¹H NMR (DMSO-d₆): δ 2.80-2.81 (d, J=4.8 Hz,3H), 7.37-7.39 (dd, J₁=2.5 Hz, J₂=5.6 Hz, 1H), 7.63-7.64 (d, J=2.6 Hz,1H), 7.67-7.74 (m, 2H), 8.03-8.05 (dd, J₁=2.8 Hz, J₂=9.36 Hz, 1H), 8.05(m, 1H), 8.12-8.14 (m, 1H), 8.14-8.16 (d, J=9.36 Hz, 1H), 8.33 (d, J=2.8Hz, 1H), 8.62-8.63 (d, J=5.7 Hz, 1H), 8.83-8.84 (q, J=4.8 Hz, 1H), 12.21(s, 1H).

Example 13 Synthesis of4-[3-(chloro-benzenesulfonylamino)-benzo[1,2,4]triazin-7-yloxy]-pyridine-2-carboxylicacid methyl amide trifluoroacetate salt

100 mg (0.337 mmol, 1.0 equivalent) of4-(3-amino-benzo[1,2,4]triazin-7-yloxy)-pyridine-carboxylic acidmethylamide were dissolved in 2 mL of anhydrous DMF with heating toabout 100° C. 45.4 mg (0.405 mmol, 1.2 equivalent) of solid t-BuOK wasadded to the solution. The resulting dark-red solution was stirred at100° C. for 30 min, then it was allowed to cool down to ambienttemperature. 85.8 mg (0.405 mmol, 1.2 equivalent) of 3-chloro-benzenesulfonyl chloride was added to the mixture via a syringe. It was allowedto stir at ambient temperature for 2 hours. The reaction mixture wasfiltered through 0.22μ syringe filter and purified by reverse-phaseprep-HPLC using acetonitrile/water mixture with 0.1% of TFA.

ESI-MS: [M+H]⁺, 471, 473, 474. ¹H NMR (DMSO-d₆): δ 2.78-2.79 (d, J=4.9Hz, 3H), 7.30-7.32 (dd, J₁=2.6 Hz, J₂=5.6 Hz, 1H), 7.57 (d, J=2.6 Hz,1H), 7.65-7.68 (t, J=9.0 Hz, 1H), 7.75-7.77 (m, 1H), 7.98-7.99 (m, 2H),8.10-8.12 (d, J=7.8 Hz, 1H), 8.15 (t, J=1.8 Hz, 1H), 8.22 (d, J=2.5 Hz,1H), 8.57-8.58 (d, J=5.4 Hz, 1H), 8.80-8.83 (q, J=4.9 Hz, 1H).

Example 14 Synthesis of4-[2-(trifluoromethyl-benzenesulfonylamino)-benzo[1,2,4]triazin-7-yloxy]-pyridine-2-carboxylicacid methyl amide trifluoroacetate salt

100 mg (0.337 mmol, 1.0 equivalent) of4-(3-amino-benzo[1,2,4]triazin-7-yloxy)-pyridine-carboxylic acidmethylamide were dissolved in 2 mL of anhydrous DMF with heating toabout 100° C. 45.4 mg (0.405 mmol, 1.2 equivalent) of solid t-BuOK wasadded to the solution. The resulting dark-red solution was stirred at100° C. for 30 min, then it was allowed to cool down to ambienttemperature. 100 mg (0.405 mmol, 1.2 equivalent) of2-trifluoromethyl-benzenesulfonyl chloride was added to the mixture viaa syringe. It was allowed to stir at ambient temperature for 2 hours.The reaction mixture was filtered through 0.22μ syringe filter andpurified by reverse-phase prep-HPLC using acetonitrile/water mixturewith 0.1% of TFA.

ESI-MS: [M+H]⁺, 505, 506, 507. ¹H NMR (DMSO-d₆): δ 2.78-2.79 (d, J=4.8Hz, 3H), 7.29-7.31 (dd, J₁=2.6 Hz, J₂=5.7 Hz, 1H), 7.56 (d, J=2.6 Hz,1H), 7.87-7.99 (m, 5H), 8.21 (d, J=2.6 Hz, 1H), 8.56-8.58 (d, 1H),8.59-8.60 (d, J=8.0 Hz, 1H), 8.80-8.81 (q, J=4.8 Hz, 1H).

Example 15 Synthesis of4-[2-chloro-5-(trifluoromethyl-benzenesulfonylamino)-benzo[1,2,4]triazin-7-yloxy]-pyridine-2-carboxylicacid methyl amide trifluoroacetate salt

The experimental procedure that was used was the same as described inExample 9.

ESI-MS: [M+H]⁺, 539, 541, 542. ¹H NMR (DMSO-d₆): δ (ppm) 2.78-2.79 (d,J=4.9 Hz, 3H), 7.27-7.29 (dd, J₁=2.6 Hz, J₂=5.7 Hz, 1H), 7.55 (d, J=2.6Hz, 1H), 7.63-7.65 (d, J=9.1 Hz, 1H), 7.86-7.88 (d, J=8.4 Hz, 1H),7.91-7.94 (dd, J₁=2.6 Hz, J₂=9.1 Hz, 1H), 8.04-8.06 (dd, J₁=2.0 Hz,J₂=8.4 Hz, 1H), 8.17-8.18 (d, J=2.6 Hz, 1H), 8.56-8.57 (d, J=5.7 Hz,1H), 8.60-8.61 (d, J=2.0 Hz, 1H), 8.80-8.81 (q, J=4.9 Hz, 1H).

Example 16 Synthesis of4-[3-chloro-6-methoxy-benzenesulfonylamino)-benzo[1,2,4]triazin-7-yloxy]-pyridine-2-carboxylicacid methyl amide trifluoroacetate salt

The experimental procedure that was used was the same as described inExample 9.

ESI-MS: [M+H]⁺, 501, 503, 504. ¹H NMR (DMSO-d₆): δ (ppm) 2.78-2.79 (d,J=4.9 Hz, 3H), 3.85 (s, 3H), 7.20-7.22 (d, J=8.9 Hz, 1H), 7.29-7.31 (dd,J=2.6 Hz, J₂=5.7 Hz, 1H), 7.55 (d, J=2.6 Hz, 1H), 7.67-7.70 (dd, J₁=2.8Hz, J₂=8.9 Hz, 1H), 7.80-7.81 (d, J=9.2 Hz, 1H), 7.92-7.95 (dd, J₁=2.8Hz, J₂=9.2 Hz, 1H), 8.05-8.06 (d, J=2.8 Hz, 1H), 8.19-8.20 (d, J=2.6 Hz,1H), 8.57-8.58 (d, J=5.6 Hz, 1H), 8.80-8.81 (q, J=4.9 Hz, 1H), 13.00(br.s. 1H).

Example 17 Synthesis of4-[3-(5-chloro-thiophene-2-sulfonylamino)-benzo[1,2,4]triazin-7-yloxy]-pyridine-2-carboxylicacid methyl amide trifluoroacetate salt

The experimental procedure that was used was the same as described inExample 9.

ESI-MS: [M+H]⁺, 477, 479. ¹H NMR (DMSO-d₆): δ (ppm) 2.79-2.80 (d, J=4.8Hz, 3H), 7.26-7.27 (d, J=4.1 Hz, 1H), 7.32-7.34 (dd, J₁=2.6 Hz, J₂=5.6Hz, 1H), 7.59 (d, J=2.6 Hz, 1H), 7.88-7.89 (d, J=4.1 Hz, 1H), 7.99-8.02(dd, J₁=2.7 Hz, J₂=9.1 Hz, 1H), 8.12-8.14 (d, J=9.1 Hz, 1H), 8.26 (d,J=2.7 Hz, 1H), 8.59 (d, J=5.6 Hz, 1H), 8.82-8.83 (q, J=4.8 Hz, 1H),13.00 (br.s. 1H).

Example 18 Synthesis of4-[2-chloro-3-trifluoromethyl-benzoylamino)-benzo[1,2,4]triazin-7-yloxy]-pyridine-2-carboxylicacid methyl amide trifluoroacetate salt

The experimental procedure that was used was the same as described inExample 10.

ESI-MS: [M+H]⁺, 503, 505, 506. ¹H NMR (DMSO-d₆): δ (ppm) 2.80-2.81 (d,J=4.8 Hz, 3H), 7.36-7.37 (dd, J₁=2.6 Hz, J₂=5.6 Hz, 1H), 7.61 (d, J=2.6Hz, 1H), 7.82-7.83 (d, J=8.4 Hz, 1H), 7.90-7.92 (dd, J₁=2.1 Hz, J₂=8.4Hz, 1H), 8.00-8.03 (m, 2H), 8.09 (m, 1H), 8.29 (m, 1H), 8.61-8.62 (d,J=5.6 Hz, 1H), 8.83-8.84 (q, J=4.8 Hz, 1H), 12.41 (s, 1H).

Example 19 Synthesis of4-[2-chloro-3-trifluoromethyl-benzoylamino)-benzo[1,2,4]triazin-7-yloxy]-pyridine-2-carboxylicacid methyl amide trifluoroacetate salt

The experimental procedure that was used was the same as described inExample 10.

ESI-MS: [M+H]⁺, 435, 437. ¹H NMR (DMSO-d₆): δ (ppm) 2.80-2.81 (d, J=4.8Hz, 3H), 7.37-7.39 (dd, J₁=2.6 Hz, J₂=5.6 Hz, 1H), 7.59-7.62 (t, J=7.8Hz, 1H), 7.63-7.64 (d, J=2.6 Hz, 1H), 7.72-7.74 (m, 1H), 8.03-8.05 (m,2H), 8.13 (m, 1H), 8.15 (d, J=9.1 Hz, 1H), 8.33 (d, J=2.6 Hz, 1H),8.62-8.63 (d, J=5.6 Hz, 1H), 8.83-8.84 (q, J=4.8 Hz, 1H), 12.12 (s, 1H).

Example 20 Synthesis of4-[2,4-dichloro-benzoylamino)-benzo[1,2,4]triazin-7-yloxy]-pyridine-2-carboxylicacid methyl amide trifluoroacetate salt

The experimental procedure that was used was the same as described inExample 10.

ESI-MS: [M+H]⁺, 469, 471, 472. ¹H NMR (DMSO-d₆): δ (ppm) 2.80-2.81 (d,J=4.8 Hz, 3H), 7.35-7.37 (dd, J₁=2.6 Hz, J₂=5.6 Hz, 1H), 7.55-7.57 (dd,J₁=1.9 Hz, J₂=8.3 Hz, 1H), 7.61 (d, J=2.6 Hz, 1H), 7.68-7.69 (d, J=8.3Hz, 1H), 7.76-7.77 (d, J=1.9 Hz, 1H), 8.01 (m, 2H), 8.28-8.29 (m, 1H),8.61 (d, J=5.6 Hz, 1H), 8.83-8.84 (q, J=4.8 Hz, 1H), 12.28 (s, 1H).

Example 21 Synthesis of4-[2-fluoro-3-chloro-5-trifluoromethyl-benzoylamino)-benzo[1,2,4]triazin-7-yloxy]-pyridine-2-carboxylicacid methyl amide trifluoroacetate salt

The experimental procedure that was used was the same as described inExample 10.

ESI-MS: [M+H]⁺, 521, 523, 524, ¹H NMR (DMSO-d₆): δ (ppm) 2.80-2.81 (d,J=4.8 Hz, 3H), 7.36-7.38 (dd, J₁=2.6 Hz, J₂=5.6 Hz, 1H), 7.62 (d, J=2.6Hz, 1H), 8.02-8.05 (dd, J₁=2.6 Hz, J₂=9.2 Hz, 1H), 8.07-8.09 (d, J=9.2Hz, 1H), 8.14-8.15 (dd, 1H), 8.31-8.32 (d, 2.6 Hz, 1H), 8.33-8.34 (dd,J₁=2.1 Hz, J₂=6.4 Hz, 1H), 8.62 (d, J=5.6 Hz, 1H), 8.83-8.84 (q, J=4.8Hz, 1H), 12.45 (s, 1H).

Example 22 Synthesis of4-[3-(4-chloro-3-trifluoromethyl-benzoylamino)-benzo[1,2,4]triazin-7-yloxy]pyridine-2-carboxylicacid meth amide trifluoroacetate salt

The experimental procedure that was used was the same as described inExample 10,

ESI-MS: [M+H]⁺, 503, 505, 506. ¹H NMR (DMSO-d₆): δ (ppm) 2.80-2.81 (d,J=4.8 Hz, 3H), 7.37-7.39 (dd, J₁=2.6 Hz, J₂=5.6 Hz, 1H), 7.63-7.64 (d,J=2.6 Hz, 1H), 7.95-7.97 (d, J=8.4 Hz, 1H), 8.04-8.06 (dd, J₁=2.7 Hz,J₂=9.3 Hz, 1H), 8.15-8.17 (d, J=9.3 Hz, 1H), 8.33-8.34 (d, J=2.7 Hz,1H), 8.34-8.36 (dd, J₁=2.1 Hz, J₂=8.4 Hz, 1H), 8.54-8.56 (d, J=2.1 Hz,1H), 8.62-8.63 (d, J=5.6 Hz, 1H), 8.83-8.84 (q, J=4.8 Hz, 1H), 12.35 (s,1H).

Example 23 Synthesis of4-[3-(2-chloro-3-trifluoromethyl-benzoylamino)-benzo[1,2,4]triazin-7-yloxy]-pyridine-2-carboxylicacid methyl amide trifluoroacetate salt

The experimental procedure that was used was the same as described inExample 10.

ESI-MS: [M+H]⁺, 503, 505, 506. ¹H NMR (DMSO-d₆): δ (ppm) 2.80-2.81 (d,J=4.8 Hz, 3H), 7.35-7.36 (dd, J₁=2.6 Hz, J₂=5.5 Hz, 1H), 7.60-7.61 (d,J=2.6 Hz, 1H), 7.66-7.69 (t, J=7.8 Hz, 1H), 7.92-7.94 (m, 2H), 7.99-8.02(m, 2H), 8.29 (d, J=2.8 Hz, 1H), 8.61-8.62 (d, J=5.6 Hz, 1H), 8.83-8.84(q, J=4.8 Hz, 1H), 12.43 (s, 1H).

Example 24 Synthesis of4-[3-(3-trifluoromethoxy-benzoylamino)-benzo[1,2,4]triazin-7-yloxy]-pyridine-2-carboxylicacid methyl amide trifluoroacetate salt

The experimental procedure that was used was the same as described inExample 10,

ESI-MS: [M+H]⁺, 485, 486, 487. ¹H NMR (DMSO-d₆): δ (ppm) 2.80-2.81 (d,J=4.8 Hz, 3H), 7.35-7.37 (dd, J₁=2.6 Hz, J₂=5.6 Hz, 1H), 7.48-7.50 (d,J=8.3 Hz, 1H), 7.53-7.56 (dt, J₁=0.9 Hz, J₂=7.6 Hz, 1H), 7.62 (d, J=2.6Hz, 1H), 7.66-7.68 (dt, J₁=1.7 Hz, J₂=8.0 Hz, 1H), 7.78-7.79 (dt, J₁=1.7Hz, J₂=7.6 Hz, 1H), 7.99-8.02 (m, 2H), 8.29 (d, J=2.6 Hz, 1H), 8.61 (d,J=5.6 Hz, 1H), 8.83-8.84 (q, J=4.8 Hz, 1H), 12.22 (s, 1H).

Example 25 Synthesis of4-{3-[(5-Methyl-isoxazole-3-carbonyl)-amino]-benzo[1,2,4]triazin-7-yloxy]-pyridine-2-carboxylicacid methyl amide trifluoroacetate salt

The experimental procedure that was used was the same as described inExample 10. ¹H NMR (DMSO-d₆): δ (ppm) 2.53 (s, 3H), 2.80-2.81 (d, J=4.8Hz, 3H), 6.79 (s, 1H), 7.37-7.38 (dd, J₁=2.5 Hz, J₂=5.5 Hz, 1H),7.63-7.64 (d, J=2.4 Hz, 1H), 8.04-8.06 (dd, J₁=2.5 Hz, J₂=9.2 Hz, 1H),8.15-8.17 (d, J=9.2 Hz, 1H), 8.32-8.33 (d, J=2.5 Hz, 1H), 8.61-8.62 (d,J=5.5 Hz, 1H), 8.83-8.84 (q, J=4.8 Hz, 1H), 12.04 (s, 1H).

Example 26 Testing of Inhibition of Raf Kinase In Vitro

The ability of compounds of general structure (A) of the presentinvention to inhibit the kinase activity of Raf1 was evaluated using twomethods: a direct and a Raf1-MEK1 assay. In the direct assay, kinasereactions were conducted in 96-well plates by combining recombinanthuman raf1 (29.4 U/well, Upstate, Lake Placid, N.Y.), ATP (3 μM), myelinbasic protein substrate (MBP, 1 mg/ml, Upstate, Lake Placid, N.Y.), andtest agents (at concentrations ranging from about 1 nM/l to about 10μM), in the presence of kinase reaction buffer. The Raf1-MEK1 assayutilized 2.9 U/well raf1 and 0.25 ug/well inactive MEK1 (MEK1 inactive,Upstate, Lake Placid, N.Y.) and 3 uM ATP. After reacting for 60 minutesat 30° C., residual ATP was measured using a luciferase-based assay(KinaseGlo, Promega Corp.) as a measure of kinase activity. Data fromfour wells were then averaged and used to determine IC₅₀ values for thetest compounds (Prism software package, GraphPad Software, San DiegoCalif.).

The test results were as follows: a known Raf inhibitor, compound A,displayed an IC₅₀ of 16 nM; a known Raf inhibitor, compound B, displayedan IC₅₀ of 43 nM; and an invention compound C, showed an IC₅₀ of 76 nM.Other invention compounds exemplified in FIG. 1, displayed an IC₅₀ below100 μM.

Example 27 Testing of Inhibition of MAPK Pathway in Cellular Assay

Western Blot: Early passage primary human umbilical vein endothelialcells (HUVECs) were maintained in EGM-2 containing SingleQuots (Cambrex,East Rutherford, N.J.), 10% FBS, 10 mM HEPES, and 50 μg/ml gentamicin.Prior to treatment of the cells with inhibitor, the HUVECs were starvedfor 18 h by replacing serum-containing complete media with scrum-freeand SingleQuot-free media. The starved cells were pre-treated withinhibitors for 60 min at various concentrations (0-20 μM). Next theHUVECs were treated with 50 ng/ml VEGF or FGF (Peprotech, Rocky Hill,N.J.) for 6 min and the cells were immediately washed with ice-cold PBS.Cells were lysed with ice-cold RIPA buffer containing 100 mM Tris pH7.5, 150 mM NaCl, 1 mM EDTA, 1% deoxycholate, 1% Triton X-100, 0.1% SDS,2 mM PMSF, one Complete-Mini protease inhibitor tablet (Roche,Indianapolis, Ind.; 1 tablet/7 ml of lysis buffer) and the phophataseinhibitors NaF (500 mM) and orthovanadate (1 mM). The cells were scrapedand lysates transferred and centrifuged at 15,000 g for 10 min.Supernatants were transferred to new tubes and protein concentration wasquantitated using the BCA protein reagent (Pierce, Rockford, Ill.). Celllysates containing 20 μg of total protein were separated by 10%SDS-PAGE, transferred to nitrocellulose, and blocked in 5% milk in TBST.Anti phospho-ERK Thr 202/Tyr 204 (Cell Signaling, Beverly, Mass.),anti-phospho-MEK Ser217/221 (Cell Signaling), and c-Raf (BD BiosciencesPharmingen, San Diego, Calif.) used as primary antibodies were detectedwith horseradish peroxidase-conjugated goat anti-mouse or rabbitsecondary antibodies and bands were visualized using the SuperSignalWest Pico chemiluminescence reagent system (Pierce) and Kodak X-ray film(Rochester, N.Y.).

Bay 43-9006 (Raf/FGF inhibitor) showed reduction of expression of p-MEKand p-ERK with IC50 between 200 and 300 mM when tested in this assay.U0126 (MEK inhibitor) showed reduction in p-Erk levels with IC₅₀ between200 and 300 nM, while p-MEK levels were unaffected. The results areshown in Table 1. As can be seen, compounds of the invention showedreduction in p-MEK and p-ERK levels with IC₅₀ between 400 nM and 20 μM.

Example 28 Cell Viability Assay

XTT assay: HUVECs were seeded at 10,000 cells/well of a tissue culturetreated 96-well plate treated with collagen type I and grown overnightin the complete EGM-2 media as described above. The following morning,the inhibitors were serial diluted with DMSO and added to the cells witha final DMSO concentration of 1%. After 24-48 hours cell viability wasmeasured with an XTT assay (Sigma, St. Louis, Mo.). The cells were alsophotographed to compare morphological differences to the XTT trendsobserved. Determination of the IC₅₀ values was performed withquantitative software (Prism software package, GraphPad Software, SanDiego Calif.). Several inhibitors blocked cell proliferation and inducedapoptosis at concentrations below 1 μM and experiments were repeatedthree times to confirm the observations. The compounds of the inventiondisplayed IC₅₀ between 100 nM and 40 uM in this assay (Table 1).

TABLE 1 Test Results for Examples 26, 27 and 28. RAF-MEK assayInhibition of HUVEC cell Structure Examples (biochemical assay) WesternBlot prolifiration (IC50)

4-[3-(4-Chloro-phenylamino)- benzo[1,2,4]triazin-7-yloxy]-pyridine-2-carboxylic acid methyl amide 10-50 uM in Raf-Mek; 77 nM in direct assay;100% window not active at 5 uM

4-[3-(4-Chloro-3-trifluoromethyl- phenylamino)-benzo[1,2,4]triazin-7-yloxy]-pyridine-2-carboxylic acid methyl amide 11.6 nM; ~20% window notactive at 5 uM

4-[3-(3-Trifluromethyl- benezenesulfonylamino)-benzo[1,2,4]triazin-7-yloxy] pyridine-2- carboxylic acid methyl amideflat not active at 5 uM

4-[3-(4-Chloro-benzenesulfonylamino)- benzo[1,2,4]triazin-7-yloxy]pyridine-2- carboxylic acid methyl amide flat not active at 5 uM

4-[3-(2-trifluromethyl- benzenesulfonylamino)-benzo[1,2,4]triazin-7-yloxy] pyridine-2- carboxylic acid methyl amideflat not active at 5 uM

4-[3-(2-chloro-5-trifluoromethyl- benzenesulfonylamino)-benzo[1,2,4]triazin-7-yloxy] pyridine-2- carboxylic acid methyl amide  50 uM not active at 5 uM

4-[3-(5-chloro-2-methoxy- benzenesulfonylamino)-benzo[1,2,4]triazin-7-yloxy] pyridine-2- carboxylic acid methylamide >50 uM not active at 5 uM

4-[3-(5-chloro-thiophene-2- sulfonylamino)-benzo[1,2,4]triazin-7- yloxy]pyridine-2-carboxylic acid methyl amide >50 uM not active at 5 uM

4-[3-(3-Trifluromethyl-benzoylamino)-benzo[1,2,4]triazin-7-yloxy]-pyridine-2- carboxylic acid methyl amide655 nM; ~70% window not active at 5 uM

4-[3-(2-Chloro-5-trifluromethyl- benzoylamino)-benzo[1,2,4]triazin-7-yloxy]-pyridine-2-carboxylic acid methyl amide not active at 5 uM

4-[3-(3-Chloro-benzoylamino)- benzo[1,2,4]triazin-7-yloxy]-pyridine-2-carboxylic acid methyl amide flat not active at 5 uM

4-[3-(2,4-Dichloro-benzoylamino)-benzo[1,2,4]triazin-7-yloxy]-pyridine-2- carboxylic acid methyl amideflat not active at 5 uM

4-[3-(3-Chloro-2-fluoro-5-trifluoromethyl-benzoylamino)-benzo[1,2,4]triazin-7- yloxy]-prridine-2-carboxylic acidmethyl amide 723 nM; ~30% window not active at 5 uM

4-[3-(4-Chloro-5-trifluromethyl- benzoylamino)-benzo[1,2,4]triazin-7-yloxy]-pyridine-2-carboxylic acid methyl amide 10-50 uM not active at 5uM

4-[3-(2-Chloro-3-trifluromethyl- benzoylamino)-benzo[1,2,4]triazin-7-yloxy]-pyridine-2-carboxylic acid methyl amide flat not active at 5 uM

4-{3-[Methyl-(4-trifluoromethoxy- benzoyl)-amino]-benzo[1,2,4]triazin-7-yloxy}-pyridine-2-carboxylic acid dimethyl amide partially active at 5uM

4-[3-(4-Trifluoromethoxy-benzoylamino)-benzo[1,2,4]triazin-7-yloxy]-pyridine-2- carboxylic acid(2-morpholin-4-yl-ethyl)-amide active at 5 uM  1.48 uM

4-{3-Methyl-(4-trifluoromethoxy- benzoyl)-amino]-benzo[1,2,4]triazin-7-yloxy}-pyridine-2-carboxylic acid (2- morpholin-4-yl-ethyl)-amidepartially active at 5 uM 3.084 uM

4-{3-Methyl-(4-trifluoromethoxy- benzoyl)-amino]-benzo[1,2,4]triazin-7-yloxy}-pyridine-2-carboxylic acid (2- morpholin-4-yl-ethyl)-amidepartially active at 5 uM

4-{3-[3-(4-Trifluoromethoxy-phenyl)-ureido]-benzo[1,2,4]triazin-7-yloxy}- pyridine-2-carboxylic acid(2-morpholin- 4-yl-ethyl)-amide not active at 5 uM

N-[7-(Pyridin-4-yloxy)- benzo[1,2,4]triazin-3-yl]-4-trifluoromethoxy-benzamide not active at 5 uM  7.38 uM

Example 29 Synthesis of S-methyl N-[4-chloro-3(trifluoromethyl)phenyl]isothiourea hydroiodide

4-Chloro-3-trifluoromethyl-phenylthiourea (5.0 g, 19.63 mmol) wasdissolved in ca. 80 mL of anhydrous MeOH and methyl iodide (2.93 g,20.61 mmol) was added via a syringe. The reaction mixture was refluxedfor 12 hours. Then it was cooled down to ambient temperature and solventwas removed in vacuo to give colorless oil (7.85 g), which was taken tothe next step without further purifications.

S-methyl N-[4-trifluoromethoxyphenyl]isothiourea hydroiodide, S-methylN-[4-hydroxy-phenyl]isothiourea hydroiodide, S-methylN-[3-hydroxy-phenyl]isothiourea hydroiodide and S-methylN-[3-(trifluoromethyl)phenyl]isothiourea hydroiodide were preparedaccording to the method of this example.

In the case of 1,2,4-triazoles three tautomeric structures can bepresent, as shown below:

Even though all three tautomeric structures can exist, all the genericstructures and all the examples having 1,2,4-triazole moiety are shownonly in one tautomeric form, such as 4H-1,2,4-triazole for simplicityand for the comparison with its direct analogues, such as examplescontaining 1,3,4-oxadiazole moiety. The prevailing tautomeric structuredepends on the substituents on the triazole moiety and on the reactionconditions. As has been shown in the literature, 1H-1,2,4-triazole isusually the most common tautomeric form, especially if an aminosubstituent is attached to the ring. Using only 4H-tautomeric form todraw the structures for the sake of simplicity, does not imply that thecompounds of the examples that follow necessarily exist in thatparticular tautomeric form. Using this approach, the IUPAC names for theexamples below are provided for 4H-tautomeric form only, however it isunderstood, that upon the elucidation of the exact tautomeric structurethe numbering of the substituents may differ from the one that isprovided.

Example 30 Synthesis of4-[5-(4-chloro-3-trifluoromethyl-phenylamino)-4H[1,2,4]triazol-3-yl]-phenol

4-hydroxybenzoic acid hydrazide (3.0 g, 19.66 mmol) and S-methylN-[4-chloro-3-(trifluoromethyl)phenyl]isothiourea hydroiodide (7.8 g,19.66 mmol) were suspended in 100 mL of anhydrous pyridine. The reactionmixture was refluxed for 18 hours under Ar atmosphere. Then it wascooled down to ambient temperature and pyridine was removed in vacuo.The resulting yellow oil was re-dissolved in a small amount of ethylacetate, loaded on a short pad of silica gel and eluted with 5:1hexane/ethyl acetate, then with 100% ethyl acetate to collect theproduct. The product was re-purified by a second silica gel columnchromatography, using ISCO system (80 g pre-packed column, 25 min run,20% to 50% EtOAc gradient in hexane). Solvent was removed in vacuo togive the title product as a white solid (2.83 g). Yield 40.4%.

ESI-MS: [M+H]⁺ 355.1, 356.8. ¹H NMR (DMSO-d₆): δ 6.89-6.91 (d, J=8.7 Hz,2H), 7.53-7.55 (d, J=8.8 Hz, 1H), 7.70-7.72 (dd, J₁=8.8 Hz, J₂=2.6 Hz,1H), 7.77-7.79 (d, J=8.7 Hz, 2H), 8.24-8.25 (d, J=2.6 Hz, 1H), 9.81 (s,1H), 9.98 (s, 1H), 13.62 (s, 1H).

Example 31 Synthesis of4-{4-[5-(4-chloro-3-trifluoromethyl-phenylamino)-4H-[1,2,4]triazol-3-yl]-phenoxy}-pyridine-2-carboxylicacid methylamide trifluoroacetic acid salt

ESI-MS: [M+H]⁺, 490, 491. ¹H NMR (DMSO-d₆): δ 2.78-2.79 (d, J=4.8 Hz,3H), 7.24-7.25 (m, 1H), 7.40-7.42 (d, J=8.7 Hz, 2H), 7.48 (s, 1H),7.56-7.58 (d, J=8.8 Hz, 1H), 7.79-7.82 (dd, J₁=2.6 Hz, J₂=8.8 Hz, 1H),8.07-8.09 (d, J=8.7 Hz, 2H), 8.24-8.25 (d, J=2.6 Hz, 1H), 8.55-8.56 (m,1H), 8.80-8.81 (m, 1H), 9.95 (s, 1H).

Example 32 Synthesis of(4-chloro-3-trifluoromethyl-phenyl)-{5-[4-(pyridin-3-yloxy)-phenyl]-4H-[1,2,4]triazol-3-yl}-aminetrifluoroacetic acid salt

4-[5-(4-chloro-3-trifluoromethyl-phenylamino)-4H[1,2,4]triazol-3-yl]-phenol(127.8 mg, 0.36 mmol) was dissolved in 3 mL of anhydrous DMF in a 5 mLmicrowave vial (Personal Chemistry). Solid potassiumbis(trimethylsilyl)amide (144.0 mg, 0.72 mmol) was added and thereaction mixture was stirred with heating at 80° C. for 15 min, then3-bromopyridine (68.3 mg, 0.432 mmol) was added, followed by anhydrousK₂CO₃ (50.0 mg, 0.36 mmol). Then the vial was capped and microwaved at250° C. for 30 min. Then the reaction mixture was diluted with ca. 1 mLof MeOH, filtered through 0.22 um syringe filter and purified byreverse-phase preparative HPLC in acetonitrile/water system with 0.01%TFA. The product was isolated as a TFA salt (15.1 mg).

ESI-MS: [M+H]⁺, 432, 433. ¹H NMR (DMSO-d₆): δ 7.19-7.21 (d, J=8.8 Hz,2H), 7.49-7.51 (dd, J₁=4.5 Hz, J₂=8.6 Hz, 1H), 7.53-7.55 (d, J=8.8 Hz,1H), 7.58-7.60 (m, 1H), 7.74-7.76 (m, 1H), 7.95-7.98 (d, J=8.8 Hz, 2H),8.21-8.21 (d, J=2.6 Hz, 1H), 8.44 (m, 1H), 8.48 (m, 1H), 9.88 (s, 1H).

Example 33 Synthesis of methyl 4-(pyridine-3-yloxy)benzoate

3-hydroxypyridine (6.17 g, 64.87 mmol) was dissolved in 100 mL ofanhydrous DMF under argon atmosphere. Solid K₂CO₃ (8.96 g, 64.87 mmol)was added, followed by neat methyl 4-fluorobenzoate (10.0 g, 64.87mmol). The reaction mixture was heated at 135° C. for 10 hrs. Theabsence of the starting material was confirmed by LC/MS. The reactionmixture was cooled down to ambient temperature and poured into ca. 500mL of H₂O. The resulting solution was extracted 3 times with ca. 150 mLof EtOAc (during the extraction small volumes of MeOH, Et₂O and brinewere added to facilitate the separation). Combined EtOAc layers werewashed twice with sat. NaHCO₃, twice with brine and dried over anhydrousNa₂SO₄. Solvent was removed in vacuo to give dark-red oil, which waspurified by silica gel chromatography using 1:1 mixture of EtOAc/Hexaneas eluent to give the title product (4.8 g, 32.3% yield) as yellowsolid.

ESI-MS: [M+H]⁺, 230, 231. ¹H NMR (DMSO-d₆): δ 3.83 (s, 3H), 7.09-7.12(d, J=8.8 Hz, 2H), 7.48-7.51 (dd, J₁=8.4 Hz, J₂=4.9 Hz, 1H), 7.58-7.61(dq, J₁=8.4 Hz, J₂=1.4 Hz, 1H), 7.96-7.99 (d, J=8.8 Hz, 2H), 8.46-8.47(m, 2H).

Example 34 Synthesis of 4-(pyridine-3-yloxy)benzohydrazide

Methyl 4-(pyridine-3-yloxy)benzoate (4.8 g, 20.94 mmol) was dissolved inca. 150 mL of EtOH and anhydrous hydrazine (4.08 g, 4.0 mL) was addedvia a syringe. The resulting yellow solution was refluxed for 24 hrs.Then solvent was removed in vacuo to give the title product (4.8 g, 100%yield) as yellow viscous oil, which upon standing slowly solidified.

ESI-MS: [M+H]⁺, 230, 231. ¹H NMR (DMSO-d₆): δ 4.13 (br s., 2H),7.06-7.09 (d, J=8.7 Hz, 2H), 7.45-7.47 (dd, J₁=8.4 Hz, J₂=4.9 Hz, 1H),7.51-7.54 (dq, J₁=8.4 Hz, J₂=1.4 Hz, 1H), 7.85-7.88 (d, J=8.8 Hz, 2H),8.42-8.43 (m, 2H), 9.74 (s, 1H).

Example 35 Synthesis of(4-chloro-3-trifluoromethyl-phenyl)-{5-[4-(pyridin-3-yloxy)-phenyl]-4H-[1,2,4]triazol-3-yl}-amine

4-(pyridine-3-yloxy)benzohydrazide (2.33 g, 10.2 mmol) was dissolved inca. 70 mL of anhydrous pyridine and S-methylN-[4-chloro-3-(trifluoromethyl)phenyl]isothiourea hydroiodide (4.04 g,10.2 mmol) was added. The reaction mixture was refluxed for 18 hrs underAr. The pyridine was removed in vacuo and the resulting residue waspurified by silica gel chromatography using EtOAc as eluent to give thetitle product (0.56 g) as a white solid.

ESI-MS: [M+H]⁺, 432, 433. ¹H NMR (DMSO-d₆): δ 7.20-7.23 (d, J=8.8 Hz,2H), 7.46-7.49 (dd, J₁=8.4 Hz, J₂=4.5 Hz, 1H), 7.54-7.57 (m, 2H),7.75-7.77 (dd, J₁=8.8 Hz, J₂=2.6 Hz, 1H), 7.97-7.99 (d, J=8.8 Hz, 2H),8.23-8.24 (d 2.6 Hz, 1H), 8.43 (m, 1H), 8.46 (d, J=2.6 Hz, 1H), 9.88 (s,1H), 13.91 (s, 1H).

Example 36 Synthesis of6-{4-[5-(4-chloro-3-trifluoromethyl-phenylamino)-4H-[1,2,4]-triazol-3-yl]-phenoxy}-pyrimidine-2,4-diamine

4-[5-(4-chloro-3-trifluoromethyl-phenylamino)-4H[1,2,4]triazol-3-yl]-phenol(1.3 g, 3.66 mmol) was dissolved in 18 mL of anhydrous dioxane in a10-20 md microwave vial (Personal Chemistry). Solid Cs₂CO₃ (1.19 g, 3.66mmol, 1.0 eq) was added and the reaction mixture was stirred withheating at 80° C. for 10 min, then 4-chloro-2,4-diaminopyrimidine (0.530g, 3.66 mmol) was added. The vial was capped and microwaved at 200° C.for 25 min. Then the reaction mixture was diluted with ca. 10 mL ofMeOH, transferred into a round-bottom flask and concentrated in vacuo toca. 20 mL. The resulting reddish solution was loaded on a short pad ofsilica gel and eluted first with 100% ethyl acetate to remove theunreacted starting material and then with 20% MeOH in EtOAc to elute theproduct. The product was further purified by ISCO system (80 gpre-packed column, 40 min method, 0% to 10% MeOH gradient in ethylacetate). Solvent was removed in vacuo to give the title product as anoff-white solid (0.785 g). Yield 46.3%.

ESI-MS: [M+H]⁺, 463, 464, 465. ¹H NMR (DMSO-d₆): δ 5.15 (s, 1H), 6.03(s, 2H), 6.31 (s, 2H), 7.25-7.27 (d, J=8.6 Hz, 2H), 7.55-7.57 (d, J=8.8Hz, 1H), 7.77-7.78 (m, 1H), 7.96-7.97 (d, J=8.8 Hz, 2H), 8.25-8.26 (d,J=2.6 Hz, 1H), 9.91 (s, 1H), 13.91 (s, 1H). Anal. Calcd for(C₁₉H₁₄ClF₃N₈O×0.4 EtOAc): C, 49.68; H, 3.48; N, 22.50. Found: C, 49.61;H, 3.55; N, 22.90.

Example 37 Synthesis of6-{4-[5-(4-chloro-3-trifluoromethyl-phenylamino)-4H-[1,2,4]triazol-3-yl]-phenoxy}-pyrimidine-2,4-diaminemethanesulfonic acid salt

6-{4-[5-(4-Chloro-3-trifluoromethyl-phenylamino)-4H-[1,2,4]triazol-3-yl]-phenoxy}-pyrimidine-2,4-diamine×0.4EtOAc complex (470.0 mg, 0.943 mmol) was dissolved in ca. 50 mL ofanhydrous methanol and methasulfonic acid (0.0612 mL, 0.943 mmol, 1.0eq.) was added. The resulting solution was stirred for 30 min. Solventwas removed in vacuo and the resulting light-yellow foam was dried at70° C. in high vacuum for 3 hrs to give the title compound as anoff-white solid (527.3 mg). Yield 100%.

ESI-MS: [M+H]⁺, 463, 464. ¹H NMR (DMSO-d₆): δ 2.36 (s, 3H), 5.40 (s,1H), 7.39-7.40 (d, J=8.6 Hz, 2H), 7.57-7.58 (d, J=8.8 Hz, 1H), 7.78 (m,1H), 7.80 (br.s., 4H), 8.02-8.03 (d, J=8.8 Hz, 2H), 8.25-8.26 (d, J=2.6Hz, 1H), 9.94 (s, 1H), 13.98 (br, s, 1H). Anal. Calcd for(C₁₉H₁₄ClF₃N₈O×1 CH₃SO₃H): C, 42.98; H, 3.25; N, 20.05. Found: C, 42.93;H, 3.62; N, 20.12.

Example 38 Synthesis of methyl4-[(2,6-diaminopyrimidin-4-yl)oxy]benzoate

Methyl 4-hydroxybenzoate (1.52 g, 10.0 mmol) was dissolved in 18 mL ofanhydrous dioxane in 10-20 mL microwave vial (Personal Chemistry) andsolid Cs₂CO₃ was added to this solution. The suspension was stirred atambient temperature for 10 min, then 4-chloro-2,6-diaminopyrimidine(1.45 g, 10.0 mmol) was added. The vial was capped and microwaved at200° C. for 40 min. Then MeOH was added to dissolve the formedsuspension to produce a clear amber solution. The solution wastransferred into a round-bottom flask and concentrated down to ca. 20mL. This solution was purified by silica gel chromatography using 100%ethyl acetate as eluent. The product was additionally re-crystallizedfrom ca. 50 mL of 4:1 mixture of EtOAc/MeOH. The product was filtered,washed with 40 mL of EtOAc, 40 mL of anhydrous Et₂O and dried in vacuoto give the title product as a white solid (0.812 g). Yield 31.2%.

ESI-MS: [M+H]⁺, 261.01. ¹H NMR (DMSO-d₆): δ 3.84 (s, 3H), 5.19 (s, 1H),6.06 (br. s, 2H), 6.37 (br.s, 2H), 7.19-7.21 (d, J=8.7 Hz, 2H),7.95-7.97 (d, J=8.7 Hz, 2H). ¹³C NMR (DMSO-d6) 52.1, 78.3, 121.0, 125.3,130.9, 157.8, 163.2, 165.7, 166.6, 169.2.

Example 39 Synthesis of 4-[(2,6-diaminopyrimidin-4-yl)oxy]benzohydrazide

Methyl 4-[(2,6-diaminopyrimidin-4-yl)oxy]benzoate (2.74 g, 10.52 mmol)was suspended in ca. 180 mL of anhydrous methanol and anhydroushydrazine (1.021 g, 1.0 mL, 31.85 mmol, 3.03 eq) was added to thissuspension. The reaction mixture was refluxed for 3 hours, then MeOH wasvery slowly distilled off till the total volume reached ca. 30 mL. Thissolution was allowed to stand at ambient temperature for 48 hrs. A whiteprecipitate slowly crystallized out. It was collected, washed with 40 mLof EtOAc, 40 mL of anhydrous Et₂O and dried in vacuo to give the titleproduct as a fine white powder (2.02 g). Yield 73.7%.

ESI-MS: [M+H]⁺, 261.12. NMR (DMSO-d₆): δ 4.54 (br.s., 2171), 5.13 (s,1H), 6.01 (br. s, 2H), 6.30 (br.s, 2H), 7.13-7.14 (d, J=8.7 Hz, 2H),7.83-7.84 (d, J=8.7 Hz, 2H), 9.75 (s, 1H). ¹³C NMR (DMSO-d6) 77.9,120.9, 128.5, 129.3, 155.8, 163.3, 165.4, 166.6, 169.7.

Example 40 Synthesis of6-{4-[5-(4-chloro-3-trifluoromethyl-phenylamino)-4H-[1,2,4]triazol-3-yl]phenoxy}-pyrimidine-2,4-diamine

4-[(2,6-diaminopyrimidin-4-yl)oxy]benzohydrazide (1.48 g, 5.68 mmol) andS-methyl N-[4-chloro-3-(trifluoromethyl)phenyl]isothiourea hydroiodide(2.33 g, 5.89 mmol) were suspended in 30 mL of anhydrous pyridine. Thereaction mixture was refluxed for 18 hours under Ar atmosphere. Theformed yellow solution was cooled down to ambient temperature andpyridine was removed in vacuo. The resulting yellow foamy solid wasre-dissolved in 50 mL of 5:1 EtOAc/MeOH; ca. 15 g of silica gel wasadded and solvent was removed in vacuo. The impregnated silica gel waspacked into ISCO column and the product was purified using ISCO system(80 g pre-packed column, 50 min run, 0% to 10% gradient of solvent B insolvent A [Solvent A—4 mL of MeOH, 4 ml, of Et₃N in 4 L of CH₂Cl₂;Solvent B—4 mL of Et₃N in 4 L of MeOH]). Solvent was removed in vacuo togive the title product as a white solid (1.33 g). Yield 50.5%.

ESI-MS: [M+H]⁺, 463, 464. ¹H NMR (DMSO-d₆): δ 5.15 (s, 1H), 6.03 (s,2H), 6.31 (s, 2H), 7.25-7.27 (d, J=8.6 Hz, 2H), 7.55-7.57 (d, J=8.8 Hz,1H), 7.77-7.78 (m, 1H), 7.96-7.97 (d 8.8 Hz, 2H), 8.25-8.26 (d, J=2.6Hz, 1H), 9.91 (s, 1H), 13.91 (s, 1H). Anal. Calcd for (C₁₉H₁₄ClF₃N₈O×0.4EtOAc): C, 49.68; H, 3.48; N, 22.50. Found: C, 49.61; H, 3.55; N, 22.90.

Example 41 Synthesis of6-{4-[5-(4-chloro-3-trifluoromethyl-phenylamino)-4H-[1,2,4]triazol-3-yl]-phenoxy}-pyridazin-3-ylaminetrifluoroacetic acid salt

4-[5-(4-chloro-3-trifluoromethyl-phenylamino)-4H[1,2,4]triazol-3-yl]-phenol(120.0 mg, 0.338 mmol) was dissolved in 3 mL of anhydrous DMF in a 5 mLmicrowave vial (Personal Chemistry). Solid potassiumbis(trimethylsilyl)amide (81.0 mg, 0.406 mmol) was added and thereaction mixture was stirred with heating at 80° C. for 15 min, then3-amino-6-chloro-pyridazine (48.2 mg, 0.372 mmol) was added, followed byanhydrous K₂CO₃ (46.7 mg, 0.338 mmol). Then the vial was capped andmicrowaved at 200° C. for 30 min. After reaction was complete, thereaction mixture was diluted with ca. 1 mL of MeOH, filtered through0.22 um syringe filter and purified by reverse-phase preparative HPLC inacetonitrile/water system with 0.01% TFA. The product was isolated as aTFA salt (18.2 mg).

ESI-MS: [M+H]⁺, 448, 449. ¹H NMR (DMSO-d₆): δ 7.42-7.43 (d, J=8.7 Hz,2H), 7.54-7.56 (d, J=9.7 Hz, 1H), 7.56-7.58 (d, J=8.8 Hz, 1H), 7.76-7.79(m, 1H), 7.78-7.80 (d, J=9.7 Hz, 1H), 8.01-8.04 (d J=8.8 Hz, 2H),8.25-8.26 (d, J=2.6 Hz, 1H), 8.49 (br.s., 2H), 9.94 (s, 1H).

Example 42 Synthesis of4-[5-(4-trifluoromethoxy-phenylamino)-4H[1,2,4]triazol-3-yl]-phenol

4-hydroxybenzoic acid hydrazide (0.643 g, 4.23 mmol) and S-methylN-[4-(trifluoromethoxy)phenyl]isothiourea hydroiodide (1.6 g, 4.23 mmol)were suspended in 10 mL of anhydrous pyridine. The reaction mixture wasrefluxed for 24 hours, during which time it changed color from yellowinto orange-red. Then it was cooled down to ambient temperature andpoured with stirring into 150 mL of ice-water. The formed white solidwas collected, washed thoroughly with water and dried in air. Theresulting residue was purified by silica gel chromatography using Iscocolumn with 10% to 100% gradient of ethyl acetate in hexane. Solvent wasremoved in vacuo to give the title product as a pinkish solid (575.2mg). Yield 40.4%.

ESI-MS: [M+H]⁺ 337, 338. ¹H NMR (DMSO-d₆): δ 6.85-6.87 (d, J=8.0 Hz,2H), 7.19-7.20 (d, J=8.0 Hz, 2H), 7.61-7.63 (d, J=8.7 Hz, 2H), 7.75-7.77(d, J=8.7 Hz, 2H), 9.39 (s, 1H), 9.92 (s, 1H), 13.42 (s, 1H).

Example 43 Synthesis of4-{4-[5-(4-trifluoromethoxy-phenylamino)-4H-[1,2,4]triazol-3-yl]-phenoxy}-pyridine-2-carboxylicacid methylamide trifluoroacetic acid salt

4-[5-(4-trifluoromethoxy-phenylamino)-4H[1,2,4]triazol-3-yl]-phenol(66.4 mg, 0,197 mmol) was dissolved in 2 mL of anhydrous DMF in a 5 mLmicrowave vial (Personal Chemistry). Solid potassiumbis(trimethylsilyl)amide (39.4 mg, 0.197 mmol) was added and thereaction mixture was stirred with heating at 80° C. for 15 min, then4-chloro-2-pyridinecarboxamide (33.7 mg, 0.197 mmol) was added, followedby anhydrous K₂CO₃ (27.3 mg, 0.197 mmol). Then the vial was capped andmicrowaved at 200° C. for 15 min. Then the reaction mixture was dilutedwith ca. 1 mL of MeOH, filtered through 0.22 um syringe filter andpurified by reverse-phase preparative HPLC in acetonitrile/water systemwith 0.01% TFA. The product was isolated as a TFA salt (46.6 mg).

ESI-MS: [M+H]⁺ 471, 472. ¹H NMR (DMSO-d₆): δ 2.78-2.79 (d, J=4.8 Hz,3H), 7.23-7.25 (dd, J₁=5.6 Hz, J₂=2.6 Hz, 1H), 7.25-7.27 (d, J=8.6 Hz,2H), 7.38-7.40 (d, J=8.6 Hz, 2H), 7.47-7.47 (d, J=2.6 Hz, 1H), 7.66-7.70(d, J=8.7 Hz, 2H), 8.08-8.11 (d, J=8.7 Hz, 2H), 8.55-8.56 (d, J=5.6 Hz,1H), 8.79-8.82 (t, J=4.8 Hz, 1H), 9.58 (s, 1H).

Example 44 Synthesis of{5-[4-(pyridin-4-yloxy)-phenyl]-4H-[1,2,4]triazol-3-yl]-(4-trifluoromethoxy-phenyl)-aminetrifluoroacetic acid salt

4-[5-(4-trifluoromethoxy-phenylamino)-4H[1,2,4]triazol-3-yl]-phenol(106.0 mg, 0.315 mmol) was dissolved in 2 mL of anhydrous DMF in a 5 mLmicrowave vial (Personal Chemistry). Solid potassiumbis(trimethylsilyl)amide (157.2 mg, 0.788 mmol) was added and thereaction mixture was stirred with heating at 80° C. for 15 min, then4-chloropyridine hydrochloride (56.7 mg, 0.378 mmol) was added, followedby anhydrous K₂CO₃ (44.0 mg, 0.315 mmol). Then the vial was capped andmicrowaved at 250° C. for 20 min. Then the reaction mixture was dilutedwith ca. 1 mL of MeOH, filtered through 0.22 um syringe filter andpurified by reverse-phase preparative HPLC in acetonitrile/water systemwith 0.01% TFA. The product was isolated as a TFA salt (66.5 mg ofoff-white solid).

ESI-MS: [M+H]⁺, 415, 416. ¹H NMR (DMSO-d₆): δ 7.26-7.27 (d, J=8.7 Hz,2H), 7.46-7.48 (m, 2H), 7.46-7.48 (d, J=7.1 Hz, 2H), 7.67-7.70 (d, J=8.7Hz, 2H), 8.13-8.16 (d, 8.7 Hz, 2H), 8.77-8.78 (d, J=7.1 Hz, 2H), 9.64(s, 1H).

Example 45 Synthesis of6-{4-[5-(4-trifluoromethoxy-phenylamino)-4H-[1,2,4]triazol-3-yl]-phenoxy}-pyridazin-3-ylaminetrifluoroacetic acid salt

4-[5-(4-trifluoromethoxy-phenylamino)-4H[1,2,4]triazol-3-yl]-phenol(112.0 mg, 0.33 mmol) was dissolved in 2 mL of anhydrous DMF in a 5 mLmicrowave vial (Personal Chemistry). Solid potassiumbis(trimethylsilyl)amide (132.8 mg, 0.66 mmol) was added and thereaction mixture was stirred with heating at 80° C. for 15 min, then3-amino-6-chloropyridazine (47.4 mg, 0.366 mmol) was added, followed byanhydrous K₂CO₃ (46.0 mg, 0.33 mmol). Then the vial was capped andmicrowaved at 250° C. for 15 min. Then the reaction mixture was dilutedwith ca. 1 mL of MeOH, filtered through 0.22 um syringe filter andpurified by reverse-phase preparative HPLC in acetonitrile/water systemwith 0.01% TFA. The product was isolated as a TFA salt (52.1 mg of browncrystalline solid).

ESI-MS: [M+H]⁺, 431. ¹H NMR (DMSO-d₆): δ 7.25-7.26 (d, J=8.7 Hz, 2H),7.39-7.41 (d, J=8.7 Hz, 2H), 7.52-7.54 (d, J=9.7 Hz, 1H), 7.67-7.68 (d,J=8.7 Hz, 2H), 7.76-7.78 (d, J=9.7 Hz, 1H), 8.03-8.06 (d, J=8.7 Hz, 2H),9.58 (s, 1H).

Example 46 Synthesis of6-{4-[5-(4-trifluoromethoxy-phenylamino)-4H-[1,2,4]triazol-3-yl]-phenoxy}-pyrimidine-2,4-diaminetrifluoroacetic acid salt

4-[5-(4-tTrifluoromethoxy-phenylamino)-4H[1,2,4]triazol-3-yl]-phenol(112.0 mg, 0.33 mmol) was dissolved in 2 mL of anhydrous DMF in a 5 mLmicrowave vial (Personal Chemistry). Solid potassiumbis(trimethylsilyl)amide (132.8 mg, 0.66 mmol) was added and thereaction mixture was stirred with heating at 80° C., for 15 min, then4-chloro-2,6-diamino-pyrimidine (53.0 mg, 0.366 mmol) was added,followed by anhydrous K₂CO₃ (46.0 mg, 0.33 mmol). Then the vial wascapped and microwaved at 250° C. for 15 min. Then the reaction mixturewas diluted with ca. 1 mL of MeOH, filtered through 0.22 um syringefilter and purified by reverse-phase preparative HPLC inacetonitrile/water system with 0.01% TEA. The product was isolated as aTFA salt (51.1 mg of beige solid).

ESI-MS: [M+H]⁺, 445, 446. ¹H NMR (DMSO-d₆): δ 5.36 (s, 1H), 7.25-7.27(br. d, J=8.0 Hz, 2H), 7.35-7.36 d, J=8.0 Hz, 2H), 7.67-7.68 (d, J=8.7Hz, 2H), 7.67 (br.s., 4H), 8.02-8.04 (d, J=8.7 Hz, 2H), 9.57 (br. s,1H).

Example 47 Synthesis of4-(5-{[3-(trifluoromethyl)phenyl]amino}-4H-1,2,4-triazol-3-yl)-phenol

S-methyl N-[3-(trifluoromethyl)phenyl]isothiourea hydroiodide (9.09 g,25.11 mmol) and 4-hydroxybenzoic acid hydrazide (3.82 g, 25.11 mmol)were suspended in ca. 50 mL of anhydrous pyridine under Ar. The mixturewas brought to reflux and refluxed under Ar for 12 hrs. Then thedark-yellow solution was cooled down to ambient temperature and pyridinewas removed in vacuo. The resulting reddish-yellow solid wasre-dissolved in ca. 50 mL of 4:1 mixture of EtOAc/MeOH, ca. 20 g ofsilica gel was added and solvent was removed in vacuo. The impregnatedsilica gel was loaded into 25 g ISCO sample cartridge and the productwas purified using ISCO system (solid method, 80 g column, 45 min, 0% to50% EtOAc gradient in hexane). Solvent was removed in vacuo to give thetitle product as a white solid (3.83 g). Yield 47.6%.

ESI-MS: [M+H]⁺, 321.09. ¹H NMR (DMSO-d₆): δ 6.89-6.90 (d, J=8.6 Hz, 2H),7.09-7.11 (d, J=7.3 Hz, 1H), 7.42-7.46 (t, J=7.9 Hz, 1H), 7.73-7.74 (d.J=7.0 Hz, 1H), 7.77-7.79 (d, J=8.6 Hz, 2H), 8.09 (s, 1H), 9.65 (s, 1H),9.97 (br. s., 1H).

Example 48 Synthesis of6-[4-(5-{[3-(trifluoromethyl)phenyl]amino}-4H-[1,2,4]triazol-3-yl)-phenoxy]-pyrimidine-2,4-diamine

4-(5-{[3-(trifluoromethyl)phenyl]amino}-4H-1,2,4-triazol-3-yl)phenol(160 mg, 0.5 mmol) was dissolved in 3 mL of anhydrous dioxane in 2-5 mLmicrowave vial (Personal Chemistry). Solid Cs₂CO₃ (163.0 mg, 0.5 mmol)was added, followed by 4-chloro-2,6,-diaminopyrimidine (79.5 mg, 0.55mmol). The vial was capped and microwaved at 200° C. for 20 min. Thenca. 3 mL of MeOH was added to dissolve the formed suspension, thesolution was transferred into a round-bottom flask and solvent wasremoved in vacuo. The residue was re-dissolved in 3 mL of DMF, filteredthrough 0.22 u syringe filter and purified by reverse phase preparativeHPLC using acetonitrile/water system with 0.01% of TFA.

The fractions containing the product were collected and partitionedbetween EtOAc and saturated aqueous NaHCO₃. Ethyl acetate layer waswashed with brine, dried over anhydrous sodium sulfate and filtered.Solvent was removed in vacuo to give the title compound as an off-whitesolid (92.2 mg).

ESI-MS: [M+H]⁺, 429.08. ¹H NMR (DMSO-d₆): δ 5.15 (s, 1H), 6.02 (s, 2H),6.30 (s, 2H), 7.12 (m, 1H), 7.25-7.27 (d, J=7.4 Hz, 2H), 7.45-7.47 (m,1H), 7.75-7.76 (m, 1H), 7.95-7.97 (d, J=8.6 Hz, 2H), 8.10 (s, 1H), 9.73(s, 1H), 13.84 (s, 1H).

Example 49 Synthesis of5-[4-(pyrimidin-5-yloxy)phenyl]-N-[3-(trifluoromethyl)phenyl]-4H-1,2,4-triazol-3-amine

4-(5-{[3-(trifluoromethyl)phenyl]amino}-4H-1,2,4-triazol-3-yl-phenol(100 mg, 0.31 mmol) was dissolved in 3 mL of anhydrous dioxane in 2-5 mLmicrowave vial (Personal Chemistry). Solid Cs₂CO₃ (203.4 mg, 0.62 mmol)was added, followed by 5-bromo-pyrimidine (100 mg, 0.62 mmol). Then 1 mLof anhydrous DMF was added, the vial was capped and microwaved at 250°C. for 30 min. Then ca. 3 mL of MeOH was added to dissolve the formedsuspension, the solution was transferred into a round-bottom flask andsolvent was removed in vacuo. The residue was re-dissolved in 3 mL ofDMF, filtered through 0.22 u syringe filter and purified by reversephase preparative HPLC using acetonitrile/water system with 0.01% ofTFA. The fractions containing the product were collected and partitionedbetween EtOAc and saturated aqueous NaHCO₃. Ethyl acetate layer waswashed with brine, dried over anhydrous sodium sulfate and filtered.Solvent was removed in vacuo to give the title compound as a light-brownfoamy solid (34.7 mg).

ESI-MS: [M+H]⁺ , 399.06. ¹H NMR (DMSO-d₆): δ 7.12 (m, 1H), 7.30-7.32 (d,J=7.4 Hz, 2H), 7.46 (m, 1H), 7.76 (m, 1H), 8.00-8.02 (d, J=8.6 Hz, 2H),8.09 (s, 1H), 8.73 (s, 2H), 9.06 (s, 1H), 9.73 (s, 1H), 13.95 (s, 1H).

Example 50 Synthesis of5-[4-(pyridin-3-yloxy)phenyl]-N-[3-trifluoromethyl)phenyl]-4H-[1,2,4]triazol-3-amine

4-(5-{[3-(trifluoromethyl)phenyl]amino}-4H-1,2,4-triazol-3-yl)phenol(100 mg, 0.31 mmol) was dissolved in 2 mL of anhydrous DMF in 2-5 mLmicrowave vial (Personal Chemistry). Solid Cs₂CO₃ (203.4 mg, 0.62 mmol)was added, followed by 3-bromopyridine (74.0 mg, 0.468 mmol). The vialwas capped and microwaved at 250° C. for 30 min. Then ea, 1 mL of MeOHwas added to dissolve the formed suspension. The resulting reddish-brownsolution was filtered through 0.22 u syringe filter and purified byreverse phase preparative HPLC using acetonitrile/water system with0.01% of TFA. The fractions, containing the product, were collected andpartitioned between EtOAc and saturated aqueous NaHCO₃. Ethyl acetatelayer was washed with brine, dried over anhydrous sodium sulfate andfiltered. Solvent was removed in vacuo to give the title compound as ayellow solid (22.4 mg).

ESI-MS: [M+H]⁺ , 398.11. ¹H NMR (DMSO-d₆): δ 7.09 (d, J=7.6 Hz, 1H),7.20-7.22 (d, J=8.6 Hz, 2H), 7.46-7.48 (m, 2H), 7.52-7.54 (m, 1H),7.75-7.76 (d, J=8.2 Hz, 1H), 7.99-8.02 (d, J=8.6 Hz, 2H), 8.09 (s, 1H),8.42-8.43 (d, J=4.5 Hz, 1H), 8.45-8.46 (d, J=2.6 Hz, 1H), 9.73 (s, 1H),13.85 (s, 1H).

Example 51 Synthesis of4-Methoxy-6-[4-(5-{[3-(trifluoromethyl)phenyl]amino}-4H-1,2,4-triazol-3-yl)phenoxyl]pyrimidin-2-amine

4-(5-{[3-(trifluoromethyl)phenyl]amino}-4H-1,2,4-triazol-3-yl)phenol(100 mg, 0.31 mmol) was dissolved in 3 mL of anhydrous dioxane in 2-5 mLmicrowave vial (Personal Chemistry). Solid Cs₂CO₃ (101.7 mg, 0.31 mmol)was added, followed by 2-amino-4-chloro-6-methoxypyrimidine (55.0 mg,0.34 mmol). The vial was capped and microwaved at 200° C. for 15 min.Then ca. 3 mL of MeOH was added to dissolve the formed suspension. Theresulting reddish-brown solution was transferred into a round-bottomflask and solvent was removed in vacuo. The residue was re-dissolved in3 mL of DMF, filtered through 0.22 u syringe filter and purified byreverse phase preparative HPLC using acetonitrile/water system with0.01% of TEA. The fractions, containing the product, were collected andpartitioned between EtOAc and saturated aqueous NaHCO₃. Ethyl acetatelayer was washed with brine, dried over anhydrous sodium sulfate andfiltered. Solvent was removed in vacuo to give the title compound as anoff-white solid (55.6 mg).

ESI-MS: [M+H]⁺ , 443.87. ¹H NMR (DMSO-d₆): δ 3.80 (s, 3H), 5.53 (s, 1H),6.70 (s, 2H), 7.11-7.12 (d, J=7.4 Hz, 1H), 7.30-7.32 (d, J=8.4 Hz, 2H),7.44-7.47 (t, J=7.6 Hz, 1H), 7.74-7.76 (d, J=8.4 Hz, 1H), 7.97-7.98 (d,J=8.4 Hz, 2H), 8.10 (s, 1H), 9.73 (s, 1H), 13.88 (s, 1H).

Example 52 Synthesis of6-[4-(5-{[3-(trifluoromethyl)phenyl]amino}-4H-[1,2,4]triazol-3-yl)-phenoxy]-pyrimidin-4-amine

4-(5-{[3-(trifluoromethyl)phenyl]amino}-4H-1,2,4-triazol-3-yl)phenol(100 mg, 0.31 mmol) was dissolved in 3 mL of anhydrous dioxane in 2-5 mLmicrowave vial (Personal Chemistry). Solid Cs₂CO₃ (101.7 mg, 0.31 mmol)was added, followed by 4-amino-6-chloro-pyrimidine (48.5 mg, 0.37 mmol).The vial was capped and microwaved at 200° C., for 5 min. Then ca. 3 mLof MeOH was added to dissolve the formed suspension. The resultingreddish-brown solution was transferred into a round-bottom flask andsolvent was removed in vacuo. The residue was re-dissolved in 3 mL ofDMF, filtered through 0.22 u syringe filter and purified by reversephase preparative HPLC using acetonitrile/water system with 0.01% ofTFA. The fractions, containing the product, were collected andpartitioned between EtOAc and saturated aqueous NaHCO₃. Ethyl acetatelayer was washed with brine, dried over anhydrous sodium sulfate andfiltered. Solvent was removed in vacuo to give the title compound as awhite solid (76.6 mg).

ESI-MS: [M+H]⁺ , 461.0. ¹H NMR (DMSO-d₆): δ 2.31 (s, 3H), 5.49 (s, 1H),6.93 (s, 2H), 7.14 (m, 1H), 7.33-7.35 (d, J=7.5 Hz, 2H), 7.45-7.48 (t,J=7.6 Hz, 1H), 7.75 (m, 1H), 8.00-8.02 (d, J=8.6 Hz, 2H), 8.10 (s, 1H),9.75 (s, 1H), 13.88 (s, 1H).

Example 53 Synthesis of2-(Methylthio)-6-[4-(5-{[3-(trifluoromethyl)phenyl]amino}-4H-1,2,4-triazol-3-yl)-phenoxy]-pyrimidin-4-amine

4-(5-{[3-(trifluoromethyl)phenyl]amino}-4H-1,2,4-triazol-3-yl)phenol(100 mg, 0.31 mmol) was dissolved in 3 mL of anhydrous dioxane in 2-5 mLmicrowave vial (Personal Chemistry). Solid Cs₂CO₃ (101.7 mg, 0.31 mmol)was added, followed by 4-amino-6-chloro-2-(methylthio)-pyrimidine (60.3mg, 0.34 mmol). The vial was capped and microwaved at 200° C. for 10min. Then ca. 3 mL of MeOH was added to dissolve the formed suspension.The resulting reddish-brown solution was transferred into a round-bottomflask and solvent was removed in vacuo. The residue was re-dissolved in3 mL of DMF, filtered through 0.22 u syringe filter and purified byreverse phase preparative HPLC using acetonitrile/water system with0.01% of TFA. The fractions, containing the product, were collected andpartitioned between EtOAc and saturated aqueous NaHCO₃. Ethyl acetatelayer was washed with brine, dried over anhydrous sodium sulfate andfiltered. Solvent was removed in vacuo to give the title compound as awhite solid (39.5 mg).

ESI-MS: [M+H]⁺, 461.0. ¹H NMR (DMSO-d₆): δ 3.80 (s, 3H), 5.53 (s, 1H),6.70 (s, 2H), 6.30 (s, 2H), (d, J=8.6 Hz, 2H).

Example 54 Synthesis of4-[5-(4-trifluoromethoxy-phenyl)-4H-[1,2,4]triazol-3-ylamino]-phenol

4-trifluoromethoxybenzoic acid hydrazide (1.1 g, 5.0 mmol) and S-methylN-[4-hydroxy-phenyl]isothiourea hydroiodide (1.55 g, 5.0 mmol) weresuspended in 10 mL of anhydrous pyridine. The reaction mixture wasrefluxed for 24 hours, during which time it changed color from yellowinto orange-red. Then it was cooled down to ambient temperature andpoured with stirring into 150 mL of ice-water. The aqueous layer wasdecanted and the resulting residue was purified by silica gelchromatography using 1:1 mixture of ethyl acetate/hexane. Solvent wasremoved in vacuo to give the title product as a pinkish-grey solid(684.0 mg). 40.6% yield.

ESI-MS: [M+H]⁺, 337, 338. ¹H NMR (DMSO-d₆): δ 7.68-6.71 (d, J=8.8 Hz,2H), 7.32-7.35 (d, J=8.8 Hz, 2H), 7.47-7.49 (d, J=8.8 Hz, 2H), 8.04-8.07(d, 8.8 Hz, 2H), 9.05 (br. s, 1H).

Example 55 Synthesis of4-{4-[5-(4-trifluoromethoxy-phenyl)-4H-[1,2,4]triazol-3-ylamino]-phenoxy}-pyridine-2-carboxylicacid methylamide trifluoroacetic acid salt

4-[5-(4-trifluoromethoxy-phenyl)-4H[1,2,4]triazol-3-ylamino]-phenol(134.5 mg, 0.4 mmol) was dissolved in 2 mL of anhydrous DMF in a 5 mLmicrowave vial (Personal Chemistry). Solid potassiumbis(trimethylsilyl)amide (120.0 mg, 0.6 mmol) was added and the reactionmixture was stirred with heating at 80° C. for 15 min, then4-chloro-2-pyridine-carboxamide (68.2 mg, 0.4 mmol) was added, followedby anhydrous K₂CO₃ (62.0 mg, 0.44 mmol). Then the vial was capped andmicrowaved at 150° C. for 30 min. Then the reaction mixture was dilutedwith ca. 1 mL of MeOH, filtered through 0.22 um syringe filter andpurified by reverse-phase preparative HPLC in acetonitrile/water systemwith 0.01% TFA. The product was isolated as a TFA salt (31.7 mg of whitesolid).

ESI-MS [M+H]⁺ 471, 472. ¹H NMR DMSO-d₆): δ 2.77-2.78 (d, J=4.8 Hz, 3H),7.13-7.15 (d, J=8.3 Hz, 2H), 7.13 (m, 1H), 7.40 (br, s, 1H), 7.51-7.53(d, J=8.3 Hz, 2H), 7.70-7.73 (d, J=8.8 Hz, 2H), 8.09-8.11 (d, J=8.8 Hz,2H), 8.48-8.49 (d, J=5.3 Hz, 1H), 8.77-8.78 (q, J=4.8 Hz, 1H), 9.56 (s,1H).

Example 56 Synthesis of3-[5-(4-chloro-3-trifluoromethyl-phenylamino)-4H[1,2,4]triazol-3-yl]-phenol

3-hydroxybenzoic acid hydrazide (2.98 g, 19.58 mmol) and S-methylN-[4-chloro-3-(trifluoromethyl)phenyl]isothiourea hydroiodide (7.78 g,19.63 mmol) were suspended in 40 mL of anhydrous pyridine. The reactionmixture was refluxed for 18 hours, during which time it changed colorfrom yellow into dark-red. Then it was cooled down to ambienttemperature and poured with stirring into 250 mL of ice-water. Theaqueous solution was decanted and the oily residue was purified bysilica gel chromatography on Isco column using 0=>50% gradient of ethylacetate in hexane. Solvent was removed in vacuo to give the titleproduct as a white solid (2.176 g). 31.3% yield.

ESI-MS: [M+H]⁺ 355.1, 356.8. ¹H NMR (DMSO-d₆): δ 6.88-6.90 (dq, J₁=7.9Hz, J₂=0.9 Hz, 1H), 7.31-7.34 (t, J=7.9 Hz, 1H), 7.35-7.39 (m, 2H),7.54-7.56 (d, J=8.8 Hz, 1H), 7.74-7.76 (dd, J₁=8.8 Hz, J₂=2.7 Hz, 1H),8.23-8.24 (d, J=2.7 Hz, 1H), 9.79 (s, 1H), 9.87 (s, 1H), 13.86 (s, 1H).

Example 57 Synthesis of4-{3-[5-(4-chloro-3-trifluoromethyl-phenylamino)-4H-[1,2,4]triazol-3-yl]-phenoxy}-pyridine-2-carboxylicacid methylamide trifluoroacetic acid salt

3-[5-(4-chloro-3-trifluoromethyl-phenylamino)-4H[1,2,4]triazol-3-yl]-phenol(100 mg, 0.282 mmol) was dissolved in 2 mL of anhydrous DMF in a 5 mLmicrowave vial (Personal Chemistry). Solid potassiumbis(trimethylsilyl)amide (140.6 mg, 0.705 mmol) was added and thereaction mixture was stirred with heating at 80° C. for 15 min, then4-chloro-2-pyridine-carboxamide (52.9 mg, 0.31 mmol) was added, followedby anhydrous K₂CO₃ (19.5 mg, 0.141 mmol). Then the vial was capped andmicrowaved at 250° C. for 20 min. Then the reaction mixture was dilutedwith ca. 1 mL of MeOH, filtered through 0.22 um syringe filter andpurified by reverse-phase preparative HPLC in acetonitrile/water systemwith 0.01% TFA. The product was isolated as a TFA salt (25.0 rug ofwhite solid).

ESI-MS: [M+H]⁺ 489, 490, 491. ¹H NMR (DMSO-d₆): δ 2.78-2.79 (d, J=4.8Hz, 3H), 7.25-7.26 (dd, J₁=2.5 Hz, J₂=5.5 Hz, 1H), 7.38 (m, 1H),7.48-7.49 (d, J=2.5 Hz, 1H), 7.55-7.57 (d, J=8.8 Hz, 1H), 7.67-7.70 (t,J=7.8 Hz, 1H), 7.73 (br. s., 1H), 7.78 (br. s., 1H), 7.91-7.93 (d, J=7.8Hz, 1H), 8.20-8.21 (d, J=2.6 Hz, 1H), 8.56-8.57 (d, j=5.5 Hz, 1H),8.80-8.81 (q, J=4.8 Hz, 1H), 9.93 (s, 1H).

Example 58 Synthesis of6-{3-[5-(4-chloro-3-trifluoromethyl-phenylamino)-4H-[1,2,4]triazol-3-yl]-phenoxy}-pyrimidine-2,4-diaminetrifluoroacetic acid salt

3-[5-(4-chloro-3-trifluoromethyl-phenylamino)-4H[1,2,4]triazol-3-yl]-phenol(100 mg, 0.282 mmol) was dissolved in 2 mL of anhydrous DMF in a 5 mLmicrowave vial. Solid potassium bis(trimethylsilyl)amide (140.6 mg,0.705 mmol) was added and the reaction mixture was stirred with heatingat 80° C. for 15 min, then 4-chloro-2,6-diamino-pyrimidine (44.8 mg,0.31 mmol) was added, followed by anhydrous K₂CO₃ (19.5 mg, 0.141 mmol).Then the vial was capped and microwaved at 250° C. for 20 min. Then thereaction mixture was diluted with ca. 1 mL of MeOH, filtered through0.22 um syringe filter and purified by reverse-phase preparative HPLC inacetonitrile/water system with 0.01% TFA. The product was isolated as aTFA salt (37.8 mg of beige solid).

ESI-MS: [M+H]⁺ 464, 465. ¹H NMR (DMSO-d₆): δ 5.40 (s, 1H), 7.33-7.35 (d,J=7.6 Hz, 1H), 7.57-7.59 (d, J=8.8 Hz, 1H), 7.62-7.65 (t, J=7.8 Hz, 1H),7.73 (br. s., 1H), 7.80 (br. m., 1H), 7.88-7.89 (d, J=7.8 Hz, 1H),8.21-8.22 (d, J=2.6 Hz, 1H), 9.96 (s, 1H).

Example 59 Synthesis of(4-chloro-3-trifluoromethyl-phenyl)-{5-[3-(pyridin-4-yloxy)-phenyl]-4H-[1,2,4]triazol-3-yl}-aminetrifluoroacetic acid salt

3-[5-(4-chloro-3-trifluoromethyl-phenylamino)-4H[1,2,4]triazol-3-yl]-phenol(100 mg, 0.282 mmol) was dissolved in 2 mL of anhydrous DMF in a 5 mLmicrowave vial. Solid potassium bis(trimethylsilyl)amide (140.6 rug,0.705 mmol) was added and the reaction mixture was stirred with heatingat 80° C. for 15 min, then 4-chloro-pyridine hydrochloride (46.5 mg,0.31 mmol) was added, followed by anhydrous K₂CO₃ (19.5 rug, 0.141mmol). Then the vial was capped and microwaved at 250° C. for 20 min.Then the reaction mixture was diluted with ca. 1 mL of MeOH, filteredthrough 0.22 um syringe filter and purified by reverse-phase preparativeHPLC in acetonitrile/water system with 0.01% TFA. The product wasisolated as a TFA salt (34.6 mg of beige solid).

ESI-MS: [M+H]⁺ 432, 433. ¹H NMR (DMSO-d₆): δ 7.43-7.45 (br. s. 1H),7.46-7.47 (d, J=7.0 Hz, 2H), 7.56-7.58 (d, J=8.8 Hz, 1H), 7.72-7.75 (t,0.1=7.8 Hz, 1H), 7.80 (br. s., 1H), 7.81 (br. s, 1H), 7.98-8.00 (d,f=7.8 Hz, 1H), 8.21-8.22 (d, J=2.6 Hz, 1H, 8.76-8.78 (d, J=7.0 Hz, 1H),9.98 (s, 1H).

Example 60 Synthesis of6-{3-[5-(4-chloro-3-trifluoromethyl-phenylamino)-4H-[1,2,4]triazol-3-yl]-phenoxy}-pyridazin-3-ylaminetrifluoroacetic acid salt

3-[5-(4-chloro-3-trifluoromethyl-phenylamino)-4H[1,2,4]triazol-3-yl]-phenol(100 mg, 0.282 mmol) was dissolved in 2 mL of anhydrous DMF in a 5 mLmicrowave vial. Solid potassium bis(trimethylsilyl)amide (140.6 mg,0.705 mmol) was added and the reaction mixture was stirred with heatingat 80° C. for 15 min, then 3-amino-6-chloro-pyridazine (40.2 mg, 0.31mmol) was added, followed by anhydrous K₂CO₃ (19.5 rug, 0.141 mmol).Then the vial was capped and microwaved at 250° C. for 20 min. Then thereaction mixture was diluted with ca. 1 mL of MeOH, filtered through0.22 um syringe filter and purified by reverse-phase preparative HPLC inacetonitrile/water system with 0.01% TFA. The product was isolated as aTFA salt (35.5 mg of light-brown solid).

ESI-MS: [M+H]⁺ 448, 449. ¹H NMR (DMSO-d₆): δ 7.38-7.39 (br. s. 1H),7.55-7.57 (d, J=9.6 Hz, 1H), 7.57-7.58 (d, J=8.6 Hz, 1H), 7.62-7.65 (t,J=7.8 Hz, 1H), 7.78 (br. s., 1H), 7.80 (br. s, 1H), 7.82-7.84 (d, J=9.6Hz, 1H), 7.86-7.88 (d, J=7.8 Hz, 1H), 8.22-8.23 (d, J=2.6 Hz, 1H), 8.52(br.s., 2H), 9.94 (s, 1H).

Example 61 Synthesis of4-(5-{[4-chloro-3-(trifluoromethyl)-phenyl]amino}-1,3,4-oxadiazol-2-yl)phenol

Mercury (II) oxide (yellow) (4.55 g, 21.0 mmol) was suspended in 60 mLof anhydrous MeOH under Ar. A bright-orange suspension was formed. Tothis suspension was added 4-hydroxybenzoic acid hydrazide (3.20 g, 21.0mmol) and 4-chloro-3-trifluoromethyl-phenylisothiocyanate (5.0 g, 21.0mmol). The reaction mixture was refluxed for 2 hours. The solvent wasremoved in vacuo. The black residue was redissolved in 100 mL of EtOAcand the resulting black suspension was filtered through a short pad ofsilica gel. The filtrate was mixed with 10 g of dry silica gel andsolvent was removed in vacuo. The impregnated silica gel was loaded onsilica gel column and the product was separated using a gradient ofhexane:ethyl acetate mixture starting from 50:50 ratio and finishing at0:100. All fractions containing the product were combined; solvent wasremoved in vacuo to give a grey solid. The solid was heated in 50 mL of4:1 mixture of EtOAc/MeOH. The formed suspension was cooled down toambient temperature and filtered to give the title product as a whitecrystalline solid (4.54 g, 60.7% yield).

ESI-MS: [M+H]⁺ 356.0. ¹H NMR (DMSO-d₆): δ 6.92-6.95 (d, J=8.8 Hz, 2H),7.69-7.70 (d, J=8.8 Hz, 1H), 7.71-7.73 (d, J=8.8 Hz, 2H), 7.82-7.84 (dd,J₁=8.8 Hz, J₂=2.6 Hz, 1H), 8.16-8.17 (d, J=2.6 Hz, 1H), 10.21 (br s.,1H), 11.11 (br s., 1H).

Example 62 Synthesis of6-[4-(5-{[4-Chloro-3-trifluoromethyl-phenyl]amino}-1,3,4-oxadiazol-2-yl)-phenoxy]-pyrimidine-2,4-diamine

4-(5-{[4-chloro-3-(trifluoromethyl-phenyl]amino}-1,3,4-oxadiazol-2-yl)phenol(1.067 g, 3.0 mmol) was dissolved in ca. 70 mL of anhydrous DMF underargon. Solid potassium bis(trimethylsilyl)amide (0.718 g, 3.6 mmol) wasadded and the resulting yellow solution was heated at 70° C. for 1.5hours. Then solid K₂CO₃ (0.414 g, 3.0 mmol) was added, followed by2,6-diamino-4-chloropyrimidine (0.520 g, 3.6 mmol). The reaction mixturewas lets to reflux under argon for 30 hours. Then it was allowed to cooldown to ambient temperature and poured into ca. 500 mL of water. Theaqueous mixture was extracted 5 times with 100 mL of EtOAc. CombinedEtOAc extracts were washed three times with 100 mL of brine, and driedover anhydrous Na₂SO₄. Solvent was removed in vacuo to a give areddish-yellow residue, which was purified by silica gel chromatographyusing EtOAc as an eluent. Fractions, containing the product, werecollected; solvent was removed in vacuo to give the product as areddish-yellow solid. The solid was re-crystallized from 10 mL of EtOAc,collected, washed thoroughly with diethyl ether and dried in vacuo togive the title compound (0.527 g, 38% yield) as beige solid.

ESI-MS: [M+H]⁺ 464, 465. ¹H NMR (DMSO-d₆): δ 5.20 (s, 1H), 6.05 (br s.,2H), 6.34 (br.s., 2H), 7.27-7.30 (d, J=8.7 Hz, 2H), 7.71-7.72 (d, J=8.8Hz, 1H), 7.84-7.88 (dd, 8.8 Hz, J₂=2.6 Hz, 1H), 7.89-7.91 (d, J=8.7 Hz,2H), 8.18-8.19 (d, J=2.6 Hz, 1H), 11.26 (s, 1H). Anal. Calcd forC₁₉H₁₃ClF₃N₇O₂: C, 49.20; H, 2.83; N, 21.14. Found: C, 49.08; H, 3.21;N, 20.95.

Example 63 Synthesis of6-[4-(5-{[4-chloro-3-trifluoromethyl-phenyl]amino}-1,3,4-oxadiazol-2-yl)-phenoxy]-pyrimidine-2,4-diaminetrifluoroacetic acid salt

4-(5-{[4-chloro-3-(trifluoromethyl)-phenyl]amino}-1,3,4-oxadiazol-2-yl)phenol(100 mg, 0.281 mmol) was dissolved in 2.5 mL of anhydrous DMF in a 5 mLmicrowave vial. Solid potassium bis(trimethylsilyl)amide (140.6 mg,0.703 mmol) was added and the reaction mixture was stirred with heatingat 80° C. for 15 min, then 6-chloro-2,4-diamino-pyrimidine (81.3 mg,0.562 mmol) was added, followed by anhydrous K₂CO₃ (19.5 mg, 0.141mmol). Then the vial was capped and microwaved at 200° C. for 15 min.Then the reaction mixture was diluted with ca. 1 mL of MeOH, filteredthrough 0.22 um syringe filter and purified by reverse-phase preparativeHPLC in acetonitrile/water system with 0.01% TFA. The product wasisolated as a TFA salt (75.8 mg of beige solid).

ESI-MS: [M+H]⁺ 464, 465. ¹H NMR (DMSO-d₆): δ 5.41 (s, 1H), 7.39-7.42 (d,J=8.7 Hz, 2H), 7.63 (br s., 4H), 7.72-7.74 (d, J=8.8 Hz, 1H), 7.85-7.87(dd, J₁=8.8 Hz, J₂=2.7 Hz, 1H), 7.95-7.97 (d, J=8.7 Hz, 2H), 8.20 (d,J=2.7 Hz, 1H), 11.29 (s, 1H).

Example 64 Synthesis ofN-[4-chloro-3-(trifluoromethyl)phenyl]-5-[4-(pyridin-3-yloxy)phenyl]-1,3,4-oxadiazol-2-aminetrifluoroacetate salt

4-(5-{[4-chloro-3-(trifluoromethyl)-phenyl]amino}-1,3,4-oxadiazol-2-yl)phenol(100 mg, 0.281 mmol) was dissolved in ca. 2 mL of anhydrous DMF underargon. Solid potassium bis(trimethylsilyl)amide (140.2 mg, 0.702 mmol)was added and the resulting yellow solution was heated at 80° C. for 15min. Then solid K₂CO₃ (19.4 mg, 0.140 mmol) was added, followed by3-bromopyridine (89.0 mg, 0.562 mmol). The reaction mixture wasmicrowaved at 250° C. for 10 min. Then it was diluted with 1 mL of MeOH,filtered and purified by preparative reverse-phase chromatography usingacetonitrile/water with 0.1% TFA gradient. The major peak having themass of the product was collected; solvent was removed in vacuo to givethe title product as a brown oil (20.6 mg).

ESI-MS: [M+H]⁺ 433.5, 434.3. ¹H NMR (DMSO-d₆): δ 7.24-7.26 (d, J=8.8 Hz,2H), 7.57-7.59 (dd, J₁=8.4 Hz, J₂=4.7 Hz, 1H), 7.68-7.71 (dq, J₁=8.4 Hz,J₂=1.4 Hz, 1H), 7.71-7.73 (d, J=8.8 Hz, ^(1H)), 7.84-7.86 (dd, J₁=8.8Hz, J₂=2.7 Hz, 1H), 7.92-7.94 (d, J=8.8 Hz, 2H), 8.17-8.18 (d, J=2.7 Hz,1H), 8.50 (br d, J=4.0 Hz, 1H), 8.54 (br s, 1H), 1.24 (s, 1H).

Example 65 Synthesis ofN-[4-chloro-3-(trifluoromethyl)phenyl]-5-[4-(pyridin-4-yloxy)phenyl]-1,3,4-oxadiazol-2-aminetrifluoroacetate salt

4-(5-{[4-chloro-3-(trifluoromethyl)-phenyl]amino}-1,3,4-oxadiazol-2-yl)phenol(100 mg, 0.281 mmol) was dissolved in ca. 2 mL of anhydrous DMF underargon. Solid potassium bis(trimethylsilyl)amide (225 mg, 1.12 mmol) wasadded and the resulting yellow solution was heated at 80° C. for 15 min.Then solid K₂CO₃ (38.8 mg, 0.281 mmol) was added, followed by4-chloropyridine hydrochloride (84.3 mg, 0.562 mmol). The reactionmixture was microwaved at 200° C. for 25 min (Initiator, Biotage). Thenit was diluted with 1 mL of MeOH, filtered through 0.22 um syringefilter and purified by preparative reverse-phase chromatography usingacetonitrile/water gradient with 0.1% TFA. The major peak having themass of the product was collected; solvent was removed in vacuo to givethe title product as a white fluffy solid (85.6 mg).

ESI-MS: [M+H]⁺ 435.3. ¹H NMR (DMSO-d₆): δ 7.37-7.38 (d, J=4.8 Hz, 2H),7.48-7.50 (d, J=8.8 Hz, 2H), 7.72-7.74 (d, J=8.8 Hz, 1H), 7.85-7.88 (dd,J=8.8 Hz, J₂=2.7 Hz, 1H), 8.03-8.06 (d, J=8.8 Hz, 1H), 8.19-8.20 (d,J=2.7 Hz, 1H), 8.74 (br s., 2H), 11.31 (s, 1H).

Example 66 Synthesis ofN-[4-chloro-3-(trifluoromethyl)phenyl]-5-[4-(pyrimidin-5-yloxy)phenyl]-1,3,4-oxadiazol-2-aminetrifluoroacetate salt

4-(5-{[4-chloro-3-(trifluoromethyl)-phenyl]amino}-1,3,4-oxadiazol-2-yl)phenol(100 mg, 0.281 mmol) was dissolved in ca. 2 mL of anhydrous DMF underargon. Solid potassium bis(trimethylsilyl)amide (140.2 mg, 0.702 mmol)was added and the resulting yellow solution was heated at 80° C. for 15min. Then solid K₂CO₃ (19.4 mg, 0.140 mmol) was added, followed by3-bromopyrimidine (89.4 mg, 0.562 mmol). The reaction mixture wasmicrowaved at 200° C. for 15 min. Then it was diluted with 1 mL of MeOH,filtered through 0.22 um syringe filter and purified by preparativereverse-phase chromatography using acetonitrile/water gradient with 0.1%TFA. The major peak having the mass of the product was collected;solvent was removed in vacuo to give the title product as a white fluffysolid (73.0 mg of white crystalline solid).

ESI-MS: [M+H]⁺ 434.3, 435.3. 1H NMR (DMSO-d₆): δ 7.31-7.33 (d, J=8.8 Hz,2H), 7.71-7.73 (d, J=8.8 Hz, 1H), 7.84-7.86 (dd, J₁=8.8 Hz, J₂=2.7 Hz,1H), 7.93-7.95 (d, J=8.8 Hz, 2H), 8.17-8.18 (d, J=2.7 Hz, 1H), 8.77 (s,2H), 9.09 (s, 1H), 11.25 (s, 1H).

Example 67 Synthesis of4-[4-(5-{[4-chloro-3-(trifluoromethyl)phenyl]amino}-1,3,4-oxadiazol-2-yl)phenoxy]-N-methylpyridine-2-carboxamidetrifluoroacetate salt

4-(5-{[4-chloro-3-(trifluoromethyl)-phenyl]amino}-1,3,4-oxadiazol-2-yl)phenol(100 mg, 0.281 mmol) was dissolved in ca. 2 mL of anhydrous DMF underargon. Solid potassium bis(trimethylsilyl)amide (140.2 mg, 0.702 mmol)was added and the resulting yellow solution was heated at 80° C. for 15min. Then solid K₂CO₃ (19.4 mg, 0.140 mmol) was added, followed by4-chloro-2-pyridine-carboxamide (52.7 mg, 0.309 mmol). The reactionmixture was microwaved at 200 C for 15 min. Then it was diluted with 1mL of MeOH, filtered through 0.22 um syringe filter and purified bypreparative reverse-phase chromatography using acetonitrile/watergradient with 0.1% TFA. The major peak having the mass of the productwas collected; solvent was removed in vacuo to give the title product asa white solid (67.5 mg).

ESI-MS: [M+H]⁺ 490.4, 491.3. ¹H NMR (DMSO-d₆): δ 2.79-2.80 (d, J=4.9 Hz,3H), 7.26-7.28 (dd, J₁=5.6 Hz, J₂=2.6 Hz, 1H), 7.43-7.44 (d, J=6.8 Hz,2H), 7.49 (d, J=2.6 Hz, 1H), 7.72-7.74 (d, J=8.8 Hz, 1H), 7.85-7.86 (dd,J=8.8 Hz, J₂=2.6 Hz, 1H), 8.01-8.03 (d, J=6.8 Hz, 2H), 8.19 (d, J=2.6Hz, 1H), 8.57-8.58 (d, J=5.7 Hz, 1H), 8.79-8.81 (q, J=4.9 Hz, 1H), 11.28(s, 1H).

Example 68 Synthesis of4-[5-(4-chloro-3-(trifluromethyl)-phenyl)-4H-1,2,4-triazol-3-ylamino]phenol

4-chloro-3-trifluoromethylbenzoic acid hydrazide (2.89 g, 12.1 mmol) andS-methyl N-(4-hydroxyphenyl)isothiourea hydroiodide (3.75 g, 12.1 mmol)were suspended in 40 mL of anhydrous pyridine. The reaction mixture wasrefluxed for 18 hours, during which time it changed color from yellowinto dark-red. Then it was cooled down to ambient temperature and pouredwith stirring into 250 mL of ice-water. The aqueous solution wasdecanted and the oily residue was purified by silica gel chromatographyusing 1:1 mixture of ethyl acetate/hexane. Solvent was removed in vacuoto give the title product as a white solid (1.95 g). Yield 45.5%.

Example 69 Synthesis of6-[4-({5-[4-chloro-3-(trifluoromethyl)phenyl-4H-1,2,4-triazol-3-yl}amino)phenoxy]pyrimidine-2,4-diaminetrifluoroacetate salt

4-[5-(4-chloro-3-(trifluoromethyl)-phenyl)-4H-1,2,4-triazol-3-ylamino]phenol(100 mg, 0.282 mmol) was dissolved in 2 mL of anhydrous DMF. Solidpotassium bis(trimethylsilyl)amide (140.6 mg, 0.705 mmol) was added andthe resulting solution was heated at 80° C. for 15 min. Then solid K₂CO₃(20 mg, 0.141 mmol) was added, followed by4-chloro-2,6-diamino-pyrimidine (61.1 mg, 0.422 mmol). The reactionmixture was microwaved at 200° C. for 20 min. Then it was diluted with 1mL of MeOH, filtered through 0.22 um syringe filter and purified bypreparative reverse-phase chromatography using acetonitrile/watergradient with 0.1% TFA. The major peak having the mass of the productwas collected, solvent was removed in vacuo to give trifluoroacetatesalt of the product as a white solid (28.6 mg).

ESI-MS: [M+H]⁺ 463.4, 464.4. ¹H NMR (DMSO-d₆): δ5.24 (s, 1H), 7.14-7.16(d, J=8.8 Hz, 2H), 7.65-7.66 (d, J=8.8 Hz, 2H), 7.81 (br s., 4H),7.88-7.90 (d, J=8.4 Hz, 1H), 8.23-8.25 (dd, J₁=8.4 Hz, J₂=1.7 Hz, 1H),8.37 (s, 1H), 9.66 (br s., 1H).

Example 70 Synthesis of5-[4-chloro-3-(trifluoromethyl)phenyl]-N-[4-(pyridine-4-yloxy)phenyl]-4H-1,2,4-triazol-3-aminetrifluoroacetate salt

4-[5-(4-chloro-3-(trifluoromethyl)-phenyl)-4H-1,2,4-triazol-3-ylamino]phenol(100 mg, 0.282 mmol) was dissolved in 2 mL of anhydrous DMF. Solidpotassium bis(trimethylsilyl)amide (196.2 mg, 0.983 mmol) was added andthe resulting solution was heated at 80° C. for 15 min. Then solid K₂CO₃(20 mg, 0.141 mmol) was added, followed by 4-chloropyridinehydrochloride (63.2 mg, 0.421 mmol). The reaction mixture was microwavedat 220° C. for 30 min. Then it was diluted with 1 mL of MeOH, filteredthrough 0.22 um syringe filter and purified by preparative reverse-phasechromatography using acetonitrile/water gradient with 0.1% TFA. Themajor peak having the mass of the product was collected, solvent wasremoved in vacuo to give trifluoroacetate salt of the product as alight-brown solid (23.5 mg).

ESI-MS: [M+H]⁺ 432, 433. ¹H NMR (DMSO-d₆): δ 7.23-7.25 (d, J=8.8 Hz,2H), 7.39-7.40 (d, J=7.2 Hz, 2H), 7.74-7.77 (d, J=8.8 Hz, 2H), 7.89-7.90(m, 1H), 8.24-8.26 (dd, J₁=8.4 Hz, J₂=2.0 Hz, 1H), 8.38 (s, 1H),8.73-8.74 (d, J=7.2 Hz, 2H), 9.75 (br s., 1H).

Example 71 Synthesis of5-[4-chloro-3-(trifluoromethyl)phenyl]-N-[4-(pyrimidin-5-yloxy)phenyl]-4H-1,2,4-triazol-3-aninetrifluoroacetate salt

4-[5-(4-chloro-3-(trifluoromethyl)-phenyl)-4H-1,2,4-triazol-3-ylamino]phenol(100 mg, 0.282 mmol) was dissolved in 2 mL of anhydrous DMF. Solidpotassium bis(trimethylsilyl)amide (84.1 mg, 0.421 mmol) was added andthe resulting solution was heated at 80° C. for 15 min. Then solid K₂CO₃(20 mg, 0.141 mmol) was added, followed by 5-bromopyrimidine (67.0 mg,0.421 mmol). The reaction mixture was microwaved at 220° C. for min.Then it was diluted with 1 mL of MeOH, filtered through 0.22 um syringefilter and purified by preparative reverse-phase chromatography usingacetonitrile/water gradient with 0.1% TFA. The major peak having themass of the product was collected; solvent was removed in vacuo to givetrifluoroacetate salt of the product as a light-brown solid (25.0 mg).

ESI-MS: [M+H]⁺ 432.9, 435. ¹H NMR (DMSO-d₆): δ 7.14-7.16 (d, J=8.9 Hz,2H), 7.63-7.67 (d, J=8.9 Hz, 2H), 7.87-7.89 (d, J=8.4 Hz, 1H), 8.22-8.24(dd, J₁=8.4 Hz, J₂=2.0 Hz, 1H), 8.36 (s, 1H), 8.54 (s, 2H), 8.93 (s,1H), 9.57 (br s., 1H).

Example 72 Synthesis of6-[4-({5-[4-chloro-3-(trifluoromethyl)phenyl]-4H-1,2,4-triazol-3-yl}amino)phenoxy]pyridazin-3-aminetrifluoroacetate salt

4-[5-(4-chloro-3-(trifluoromethyl)-phenyl)-4H-1,2,4-triazol-3-ylamino]phenol(100 mg, 0.282 mmol) was dissolved in 2 mL of anhydrous DMF. Solidpotassium bis(trimethylsilyl)amide (140.6 mg, 0.705 mmol) was added andthe resulting solution was heated at 80° C. for 15 min. Then solid K₂CO₃(20 mg, 0.141 mmol) was added, followed by 3-amino-6-chloropyridazine(54.6 mg, 0.421 mmol). The reaction mixture was microwaved at 220° C.for 40 min. Then it was diluted with 1 mL of MeOH, filtered through 0.22um syringe filter and purified by preparative reverse-phasechromatography using acetonitrile/water gradient with 0.1% TFA. Themajor peak having the mass of the product was collected; solvent wasremoved in vacuo to give trifluoroacetate salt of the product as alight-brown solid (23.1 mg).

ESI-MS: [M+H]⁺ 448, 449. ¹H NMR (DMSO-d₆): δ 7.16-7.18 (d, J=8.9 Hz,2H), 7.51-7.53 (d, J=9.7 Hz, 1H), 7.62-7.66 (d, J=8.9 Hz, 2H), 7.73-7.75(d, J=9.7 Hz, 1H), 7.88 (d, J=8.4 Hz, 1H), 8.23-8.25 (dd, J₁=8.4 Hz,J₂=2.0 Hz, 1H), 8.36 (s, 1H), 8.48 (br s., 2H), 9.62 (br s., 1H).

Example 73 Synthesis of4-[4-({5-[4-chloro-3-(trifluoromethyl)phenyl]-4H-1,2,4-triazol-3-yl}amino)phenoxy]-N-methylpyridine-2-carboxamidetrifluoroacetate salt

4-[5-(4-chloro-3-(trifluoromethyl)-phenyl)-4H-1,2,4-triazol-3-ylamino]phenol(100 mg, 0.282 mmol) was dissolved in 2 mL of anhydrous DMF. Solidpotassium bis(trimethylsilyl)amide (140.6 mg, 0.705 mmol) was added andthe resulting solution was heated at 80° C. for 15 min. Then solid K₂CO₃(20 mg, 0.141 mmol) was added, followed by4-chloro-2-pyridine-carboxamide (71.9 mg, 0.421 mmol). The reactionmixture was microwaved at 220° C. for 20 min. Then it was diluted with 1mL of MeOH, filtered through 0.22 um syringe filter and purified bypreparative reverse-phase chromatography using acetonitrile/watergradient with 0.1% TFA. The major peak having the mass of the productwas collected; solvent was removed in vacuo to give trifluoroacetatesalt of the product as yellow solid (25.4 mg).

ESI-MS: [M+H]⁺ 489, 490. ¹H NMR (DMSO-d₆): δ 2.77-2.78 (d, J=4.8 Hz,3H), 7.14-7.15 (dd, J₁=5.6 Hz, J₂=2.5 Hz, 1H), 7.15-7.17 (d, J=9.0 Hz,2H), 7.40 (d, J=2.5 Hz, 1H), 7.68-7.71 (d, J=9.0 Hz, 2H), 7.87-7.89 (d,J=8.4 Hz, 1H), 8.23-8.25 (dd, J₁=8.4 Hz, J₂=2.0 Hz, 1H), 8.37 (s, 1H),8.49-8.50 (d, J=5.6 Hz, 1H), 8.75-8.78 (q, J=4.8 Hz, 1H), 9.65 (br s.,1H).

Example 74 Synthesis of5-[4-chloro-3-(trifluoromethyl)phenyl]-N-[4-(pyridine-3-yloxy)phenyl]-4H-1,2,4-triazol-3-aminetrifluoroacetate salt

4-[5-(4-chloro-3-(trifluoromethyl)-phenyl)-4H-1,2,4-triazol-3-ylamino]phenol(200 mg, 0.562 mmol) was dissolved in 2 mL of anhydrous DMF. Solidpotassium bis(trimethylsilyl)amide (280.3 mg, 1.405 mmol) was added andthe resulting solution was heated at 80° C. for 15 min. Then solid K₂CO₃(40 mg, 0.282 mmol) was added, followed by 3-bromopyridine (177.6 mg,1.12 mmol). The reaction mixture was microwaved at 250° C. for 20 min.Then it was diluted with 1 mL of MeOH, filtered through 0.22 um syringefilter and purified by preparative reverse-phase chromatography usingacetonitrile/water gradient with 0.1% TFA. The major peak having themass of the product was collected, solvent was removed in vacuo to givetrifluoroacetate salt of the product as a yellow solid (17.0 mg).

ESI-MS: [M+H]⁺ 432, 433. ¹H NMR (DMSO-d₆): δ 7.09-7.11 (d, J=9.0 Hz,2H), 7.50-7.52 (m, 2H), 7.63-7.66 (d, J=9.0 Hz, 2H), 7.87-7.89 (d, J=8.4Hz, 1H), 8.22-8.25 (dd, J₁=8.4 Hz, J₂=2.0 Hz, 1H), 8.37-8.38 (m, 2H),8.43 (min, 1H), 9.56 (br s., 1H).

Example 75 Synthesis of4-(5-{[3-Trifluoromethyl)phenyl]amino}-1,3,4-oxadiazol-2-yl)phenol

Mercury (II) oxide yellow (5.33 g, 24.60 mmol) was suspended in ca. 70mL of anhydrous methanol, 4-Hydroxybenzoic acid hydrazide (3.74 g, 24.60nm mmol) was added to this bright-orange suspension, followed by3-trifluoromethylphenylisothiocyanate (5.0 g, 24.60 mmol). The reactionmixture was brought to reflux and refluxed for 2 hours. The reactionmixture turned pitch-black in color and formed black precipitate. Thenit was cooled down to ambient temperature and filtered through a shortpad of Celite, then through a short pad of silica gel. Then methanol wasremoved in vacuo and the resulting grey precipitate was re-crystallizedfrom ca. 100 mL of EtOAc. The formed white crystalline solid wasfiltered, washed with a small amount of EtOAc and dried in vacuo to givethe title product as white crystals (7.182 g). Yield 71.3%.

ESI-MS: [M+H]⁺ 322.0. ¹H NMR (DMSO-d₆): δ 6.92-6.95 (d, J=8.7 Hz, 2H),7.33-7.35 (d, J=: 8.3 Hz, 1H), 7.57-7.60 (t, J=8.0 Hz, 1H), 7.72-7.75(d, J=8.7 Hz, 2H), 7.80-7.82 (dd, J₁=8.0 Hz, J₂=1.8 Hz, 1H), 8.06 (s,1H), 10.21 (s, 1H), 10.99 (br s., 1H). ¹³C NMR (DMSO-d6) 112.9, 114.5,116.1, 117.9, 120.6, 127.6, 129.7, 130.0, 130.3, 139.6, 158.3, 158.9,160.1.

Example 76 Synthesis of6-[4-(5-{[3-(trifluoromethyl)phenyl]amino}-1,3,4-oxadiazol-2-yl)phenoxy}pyrimidine-2,4-diamine

4-(5-{[3-(trifluoromethyl)-phenyl]amino}-1,3,4-oxadiazol-2-yl)phenol(160.6 mg, 0.5 mmol) was dissolved in 3 mL of anhydrous DMF in a 2-5 mLmicrowave vial (Personal Chemistry). Solid potassiumbis(trimethylsilyl)amide (119.7 mg, 0.6 mmol) was added and the reactionmixture was stirred with heating at 80° C. for 10 min, then6-chloro-2,4-diamino-pyrimidine (86.7 mg, 0.6 mmol) was added, followedby anhydrous K₂CO₃ (69.1 mg, 0.5 mmol). Then the vial was capped andmicrowaved at 200° C. for 20 min. Then the reaction mixture was dilutedwith ca. 1 mL of MeOH, filtered through 0.22 um syringe filter andpurified by reverse-phase preparative HPLC in acetonitrile/water systemwith 0.01% TFA. Fractions, containing the product, were partitionedbetween ca. 40 mL of EtOAc and ca. 40 mL of saturated NaHCO₃. EtOAclayer was washed with brine, dried over anhydrous Na₂SO₄ and filtered.Solvent was removed in vacuo to give the title product as a beige solid(81.0 mg). Yield 37.7%.

ESI-MS: [M+H]⁺ 430.29. ¹H NMR (DMSO-d₆): δ 5.19 (s, 1H), 6.04 (s, 2H),6.33 (s, 2H), 7.27-7.30 (d, J=8.7 Hz, 2H), 7.36-7.37 (d, J=7.9 Hz, 1H),7.59-7.63 (t, J=8.0 Hz, 1H), 7.82-7.83 (dd, J₁=8.0 Hz, J₂=1.8 Hz, 1H),7.89-7.92 (d, J=8.7 Hz, 2H), 8.08 (s, 1H). 11.11 (s, 1H).

Example 77 Synthesis of5-[4-(Pyrimidin-5-yloxy)phenyl]-N-[3-(trifluoromethyl)phenyl]-1,3,4-oxadiazol-2-amine

4-(5-{[3-(trifluoromethyl)-phenyl]amino}-1,3,4-oxadiazol-2-yl)phenol(160.6 mg, 0.5 mmol) was dissolved in 3 mL of anhydrous DMF in a 2-5 mLmicrowave vial (Personal Chemistry). Solid potassiumbis(trimethylsilysilyl)amide (149.6 mg, 0.75 mmol) was added and thereaction mixture was stirred with heating at 80° C. for 10 min, then5-bromopyrimidine (119.2 mg, 0.75 mmol) was added, followed by anhydrousK₂CO₃ (69.1 mg, 0.5 mmol). Then the vial was capped and microwaved at200° C. for 20 min. Then the reaction mixture was diluted with ca. 1 mLof MeOH, filtered through 0.22 um syringe filter and purified byreverse-phase preparative HPLC in acetonitrile/water system with 0.01%TFA. Fractions, containing the product, were partitioned between ca, 40mL of EtOAc and ca. 40 mL of saturated NaHCO₃. EtOAc layer was washedwith brine, dried over anhydrous Na₂SO₄ and filtered. Solvent wasremoved in vacuo to give the title product as a beige solid (87.0 mg).Yield 43.5%.

ESI-MS: [M+H]⁺ 400.16. ¹H NMR (DMSO-d₆): δ 7.31-7.33 (d, J=8.7 Hz, 2H),7.35-7.37 (d, J=7.9 Hz, 1H), 7.59-7.62 (t, J=8.0 Hz, 1H), 7.81-7.83 (dd,J₁=8.0 Hz, J₂=1.8 Hz, 1H), 7.93-7.95 (d, J=8.7 Hz, 2H), 8.07 (s, 1H),8.76 (s, 2H), 9.08 (s, 1H), 11.11 (s, 1H).

Example 78 Synthesis of4-{5-[(4-chlorophenyl)amino]-1,3,4-oxadiazol-2-yl}phenol

Mercury (II) oxide yellow (6.38 g, 29.47 mmol) was suspended in ca. 70mL of anhydrous methanol. 4-Hydroxybenzoic acid hydrazide (4.48 g, 29.47mmol) was added to this bright-orange suspension, followed by4-chlorophenylisothiocyanate (5.0 g, 29.47 mmol). The reaction mixturewas brought to reflux and refluxed for 2 hours. The reaction mixtureturned pitch-black in color and formed black precipitate. Then it wascooled down to ambient temperature and filtered through a short pad ofCelite, then through a short pad of silica gel. Then methanol wasremoved in vacuo and the resulting grey precipitate was re-crystallizedfrom ca. 40 mL of EtOAc. The formed white precipitate was filtered,washed with a small amount of EtOAc and dried in vacuo to give the titleproduct as a white powder.

ESI-MS: [M−H]⁺ 287.94. ¹H NMR (DMSO-d₆): δ 6.91-6.94 (d, J=8.7 Hz, 2H),7.39-7.41 (d, J=8.9 Hz, 2H), 7.61-7.63 (d, J=8.9 Hz, 2H), 7.71-7.74 (d,J=8.7 Hz, 2H), 10.19 (s, 1H), 10.73 (s, 1H). ¹³C NMR (DMSO-d6) 114.6,116.1, 118.5, 125.3, 127.5, 128.9, 137.8, 158.1, 159.1, 160.0.

Example 79 Synthesis of6-(4-{5-[(4-chlorophenyl)amino]-1,3,4-oxadiazol-2-yl}phenoxy)pyrimidine-2,4-diamine

4-{5-[(4-chlorophenyl)amino]-1,3,4-oxadiazol-2-yl}phenol (144.0 mg, 0.5mmol) was dissolved in 3 mL of anhydrous DMF in a 2-5 mL microwave vial(Personal Chemistry). Solid potassium bis(trimethylsilyl)amide (100.0mg, 0.5 mmol) was added and the reaction mixture was stirred withheating at 80° C. for 10 min, then 6-chloro-2,4-diamino-pyrimidine (72.3mg, 0.5 mmol) was added, followed by anhydrous K₂CO₃ (34.5 mg, 0.25mmol). Then the vial was capped and microwaved at 200° C. for 15 min.Then the reaction mixture was diluted with ca. 1 mL of MeOH, filteredthrough 0.22 um syringe filter and purified by reverse-phase preparativeHPLC in acetonitrile/water system with 0.01% TFA. Fractions, containingthe product, were partitioned between ca. 40 mL of EtOAc and ca. 40 mLof saturated NaHCO₃. EtOAc layer was washed with brine, dried overanhydrous Na₂SO₄ and filtered. Solvent was removed in vacuo to give thetitle product as a beige solid (24.5 mg).

ESI-MS: [M+H]⁺ 396.25. ¹H NMR (DMSO-d₆): δ 5.19 (s, 1H), 6.04 (s, 2H),6.33 (s, 2H), 7.26-7.29 (d, J=8.7 Hz, 2H), 7.41-7.43 (d, J=8.9 Hz, 2H),7.63-7.65 (d, J=8.9 Hz, 2H), 7.88-7.90 (d, J=8.7 Hz, 2H), 10.85 (s,1.1).

Example 80 Synthesis of5-[4-(Pyrimidin-5-yloxy)phenyl]-N-[4-chloro-phenyl]-1,3,4-oxadiazol-2-amine

4-{5-[(4-chlorophenyl)amino]-1,3,4-oxadiazol-2-yl}phenol (144.0 mg, 0.5mmol) was dissolved in 3 mL of anhydrous DMF in a 2-5 mL microwave vial(Personal Chemistry). Solid potassium bis(trimethylsilyl)amide (100.0mg, 0.5 mmol) was added and the reaction mixture was stirred withheating at 80° C. for 10 min, then 5-bromopyrimidine (79.5 mg, 0.5 mmol)was added, followed by anhydrous K₂CO₃ (34.5 mg, 0.25 mmol). Then thevial was capped and microwaved at 200° C. for 15 min. Then the reactionmixture was diluted with ca. 1 mL of MeOH, filtered through 0.22 umsyringe filter and purified by reverse-phase preparative HPLC inacetonitrile/water system with 0.01% TFA. Fractions, containing theproduct, were partitioned between ca. 40 mL of EtOAc and ca. 40 mL ofsaturated NaHCO₃. EtOAc layer was washed with brine, dried overanhydrous Na₂SO₄ and filtered. Solvent was removed in vacuo to give thetitle product as a beige solid (61.6 mg).

ESI-MS: [M+H]⁺ 366.24. ¹H NMR (DMSO-d₆): δ 7.30-7.32 (d, J=8.7 Hz, 2H),7.41-7.43 (d, J=8.9 Hz, 2H), 7.63-7.65 (d, J=8.9 Hz, 2H), 7.92-7.94 (d,J=8.7 Hz, 2H), 8.76 (s, 2H), 9.08 (s, 1H), 10.85 (s, 1H).

Example 81 Synthesis of4-(5-{[2-chloro-5-(trifluoromethyl)phenyl]amino}-1,3,4-oxadiazol-2-yl)phenol

Mercury (II) oxide yellow (1.82 g, 8.41 mmol) was suspended in ca. 50 mLof anhydrous methanol. 4-Hydroxybenzoic acid hydrazide (1.28 g, 8.41mmol) was added to this bright-orange suspension, followed by2-chloro-5-trifluoromethyl-phenylisothiocyanate (2.0 g, 8.41 mmol). Thereaction mixture was brought to reflux and refluxed for 2 hours. Thereaction mixture turned pitch-black in color and formed blackprecipitate. Then it was cooled down to ambient temperature and filteredthrough a short pad of Celite, then through a short pad of silica gel.Then methanol was removed in vacuo and the resulting grey solid wasre-crystallized from ca. 20 mL of EtOAc. The formed white precipitatewas filtered, washed with anhydrous Et₂O and dried in vacuo to give thetitle product as a white solid (2.638 g). Yield 88.1%

ESI-MS: [M+H]⁺ 356.22. ¹H NMR (DMSO-d₆): δ 6.93-6.96 (d, J=8.7 Hz, 2H),7.41-7.43 (dd, J₁=8.3 Hz, J₂=1.7 Hz, 1H), 7.72-7.75 (m, 3H), 8.61 (s,1H), 10.23 (s, 1H), 10.30 (s, 1H).

Example 82 Synthesis of6-(4-{5-[(2-chloro-5-trifluoromethyl-phenyl)amino]-1,3,4-oxadiazol-2-yl}phenoxy)pyrimidine-2,4-diamine

4-(5-{[2-chloro-5-(trifluoromethyl)phenyl]amino}-1,3,4-oxadiazol-2-yl)phenol(177.85 mg, 0.5 mmol) was dissolved in 3 mL of anhydrous DMF in a 2-5 mLmicrowave vial (Personal Chemistry). Solid potassiumbis(trimethylsilyl)amide (200.0 mg, 1.0 mmol) was added and the reactionmixture was stirred with heating at 80° C. for 10 min, then6-chloro-2,4-diamino-pyrimidine (86.7 mg, 0.6 mmol) was added, followedby anhydrous K₂CO₃ (69.1 mg, 0.5 mmol). Then the vial was capped andmicrowaved at 180° C. for 30 min. Then the reaction mixture was dilutedwith ca. 1 mL of MeOH, filtered through 0.22 um syringe filter andpurified by reverse-phase preparative HPLC in acetonitrile/water systemwith 0.01% TFA. Fractions, containing the product, were partitionedbetween ca. 50 mL of EtOAc and ca. 50 mL of saturated aqueous NaHCO₃.EtOAc layer was washed with brine, dried over anhydrous Na₂SO₄ andfiltered. Solvent was removed in vacuo to give the title product as anoff-white solid (131.1 mg). Yield 56.5%.

ESI-MS: [M+H]⁺ 464.21. ¹H NMR (DMSO-d₆): δ 5.20 (s, 1H), 6.05 (s, 2H),6.34 (s, 2H), 7.28-7.31 (d, J=8.7 Hz, 2H), 7.46-7.48 (dd, J₁=8.3 Hz,J₂=1.8 Hz, 1H), 7.76-7.78 (d, J=8.3 Hz, 1H), 7.90-7.92 (d, J=8.7 Hz,2H), 8.62 (s, 1H), 10.47 (s, 1H).

Example 83 Synthesis of5-[4-(Pyrimidin-5-yloxy)phenyl]-N-[2-chloro-5-(trifluoromethyl)-phenyl]-1,3,4-oxadiazol-2-amine

4-(5-{[2-chloro-5-(trifluoromethyl)phenyl]amino}-1,3,4-oxadiazol-2-yl)phenol(177.85 mg, 0.5 mmol) was dissolved in 3 mL of anhydrous DMF in a 2-5 mLmicrowave vial (Personal Chemistry). Solid potassiumbis(trimethylsilyl)amide (200.0 mg, 1.0 mmol) was added and the reactionmixture was stirred with heating at 80° C. for 10 min, then5-bromopyrimidine (95.4 mg, 0.6 mmol) was added, followed by anhydrousK₂CO₃ (69.1 mg, 0.5 mmol). Then the vial was capped and microwaved at180° C. for 40 min. Then the reaction mixture was diluted with ca. 1 mLof MeOH, filtered through 0.22 um syringe filter and purified byreverse-phase preparative HPLC in acetonitrile/water system with 0.01%TFA. Fractions, containing the product, were partitioned between ca. 50mL of EtOAc and ca. 50 mL of saturated aqueous NaHCO₃. EtOAc layer waswashed with brine, dried over anhydrous Na₂SO₄ and filtered. Solvent wasremoved in vacuo to give the title product as a beige solid (129.1 mg).Yield 59.5%.

ESI-MS: [M+H]⁺ 434.20. ¹H NMR (DMSO-d₆): δ 7.32-7.35 (d, J=8.7 Hz, 2H),7.46-7.48 (dd, J₁=8.3 Hz, J₂=1.8 Hz, 1H), 7.76-7.78 (d, J=8.3 Hz, 1H),7.94-7.96 (d, J=8.7 Hz, 2H), 8.61 (s, 1H), 8.77 (s, 2H), 9.09 (s, 1H),10.47 (s, 1H).

Example 84 Synthesis of4-{5-[(3-chlorophenyl)amino]-1,3,4-oxadiazol-2-yl}-phenol

Mercury (II) oxide yellow (6.38 g, 29.47 mmol) was suspended in ca. 100mL of anhydrous methanol. 4-Hydroxybenzoic acid hydrazide (4.48 g, 29.47mmol) was added to this bright-orange suspension, followed by3-chlorophenylisothiocyanate (5.0 g, 29.47 mmol). The reaction mixturewas brought to reflux and refluxed for 2 hours. The reaction mixtureturned pitch-black in color and formed black precipitate. Then it wascooled down to ambient temperature and filtered through a short pad ofCelite. Then it was purified by silica gel chromatography using 0% to20% methanol gradient in EtOAc. Solvent was removed in vacuo and theresulting grey precipitate was re-crystallized from ca. 50 mL of EtOAc.The formed white crystalline solid was filtered, washed with a smallamount of EtOAc, anhydrous Et₂O and dried in vacuo to give the titleproduct as a white powder (7.606 g). Yield 89.7%.

ESI-MS: [M+H]⁺ 288.26. ¹H NMR (DMSO-d₆): δ 6.92-6.94 (d, J=8.7 Hz, 2H),7.00-7.02 (dd, J₁=7.9 Hz, J₂=1.8 Hz, 1H), 7.33-7.36 (t, J=8.1 Hz, 1H),7.47-7.49 (dd, J₁=8.1 Hz, J₂=1.8 Hz, 1H), 7.72-7.74 (d, J=8.7 Hz, 2H),7.76-7.78 (t, J=2.1 Hz, 1H), 10.19 (s, 1H), 10.81 (s, 1H). ¹³C NMR(DMSO-d6) 114.6, 115.5, 116.1, 116.4, 121.3, 127.6, 130.6, 133.5, 140.3,158.3, 159.0, 160.1.

Example 85 Synthesis of6-(4-{5-[(3-chlorophenyl)amino]-1,3,4-oxadiazol-2-yl}phenoxy)pyrimidine-2,4-diamine

4-{5-[(3-chlorophenyl)amino]-1,3,4-oxadiazol-2-yl}phenol (143.8 mg, 0.5mmol) was dissolved in 3 mL of anhydrous DMF in a 2-5 mL microwave vial(Personal Chemistry). Solid potassium bis(trimethylsilyl)amide (200.0mg, 1.0 mmol) was added and the reaction mixture was stirred withheating at 80° C. for 10 min, then 6-chloro-2,4-diamino-pyrimidine (86.7mg, 0.6 mmol) was added, followed by anhydrous K₂CO₃ (69.1 mg, 0.5mmol). Then the vial was capped and microwaved at 180° C. for 40 min.Then the reaction mixture was diluted with ca. 1 mL of MeOH, filteredthrough 0.22 um syringe filter and purified by reverse-phase preparativeHPLC in acetonitrile/water system with 0.01% TFA. Fractions, containingthe product, were partitioned between ca. 50 nm, of EtOAc and ca. 50 mLof saturated aqueous NaHCO₃. EtOAc layer was washed with brine, driedover anhydrous Na₂SO₄ and filtered. Solvent was removed in vacuo to givethe title product as a light-yellow solid (62.0 mg). Yield 31.3%.

ESI-MS: [M+H]⁺ 396.22. ¹H NMR (DMSO-d₆): δ 5.19 (s, 1H), 6.04 (s, 2H),6.33 (s, 2H), 7.06-7.08 (m, 1H), 7.27-7.29 (d, J=8.7 Hz, 2H), 7.37-7.41(t, J=8.1 Hz, 1H), 7.50-7.52 (dd, J₁=8.1 Hz, J₂=1.8 Hz, 1H), 7.78-7.79(t, J=2.1 Hz, 1H), 7.89-7.90 (d, J=8.7 Hz, 2H), 10.95 (s, 1H).

Example 86 Synthesis of5-[4-(Pyrimidin-5-yloxy)phenyl]-N-[3-chloro-phenyl]-1,3,4-oxadiazol-2-amine

4-{5-[(3-chlorophenyl)amino]-1,3,4-oxadiazol-2-yl}phenol (143.8 mg, 0.5mmol) was dissolved in 3 mL of anhydrous DMF in a 2-5 mL microwave vial(Personal Chemistry). Solid potassium bis(trimethylsilyl)amide (200.0mg, 1.0 mmol) was added and the reaction mixture was stirred withheating at 80° C. for 10 min, then 5-bromropyrimidine (95.4 mg, 0.6mmol) was added, followed by anhydrous K₂CO₃ (69.1 mg, 0.5 mmol). Thenthe vial was capped and microwaved at 180° C. for 30 min. Then thereaction mixture was diluted with ca. 1 mL of MeOH, filtered through0.22 um syringe filter and purified by reverse-phase preparative HPLC inacetonitrile/water system with 0.01% TFA. Fractions, containing theproduct, were partitioned between ca. 50 mL of EtOAc and ca. 50 mL ofsaturated aqueous NaHCO₃. EtOAc layer was washed with brine, dried overanhydrous Na₂SO₄ and filtered. Solvent was removed in vacuo to give thetitle product as a light-yellow solid (74.7 mg). Yield 40.8%.

ESI-MS: [M+H]⁺ 366.23. ¹H NMR (DMSO-d₆): δ 7.06-7.08 (m, 1H), 7.31-7.33(d, J=8.7 Hz, 2H), 7.37-7.41 (t, J=8.1 Hz, 1H), 7.50-7.52 (dd, J₁=8.1Hz, J₂=1.8 Hz, 1H), 7.77-7.79 (t, J=2.0 Hz, 1H), 7.93-7.94 (d, J=8.7 Hz,2H), 8.77 (s, 2H), 9.09 (s, 1H), 10.96 (s, 1H).

Example 87 Synthesis of5-[4-(Pyridin-3-yloxy)phenyl]-1,3,4-oxadiazol-2-amine hydrobromide salt

4-(pyridine-3-yloxy)benzohydrazide (3.7 g, 16.14 mmol) was dissolved in100 mL of anhydrous THF and 3.0 M solution of cyanogen bromide (5.38 mL,16.14 mmol) was added via syringe. Within 5-10 min of stirring an orangeprecipitate started to form. The reaction mixture was brought to refluxand refluxed for 1 hr. Then it was cooled down to ambient temperatureand filtered. The collected orange precipitate was washed with ca. 100mL of THF, ca. 100 mL of EtOAc, anhydrous Et2O and dried in vacuo togive the title product as an orange solid (4.40 g). Yield 81.4%.

ESI-MS: [M+H]⁺ 255.05. ¹H NMR (DMSO-d₆): δ 7.27-7.29 (d, J=8.8 Hz, 2H),7.75-7.78 (dd, J₁=8.5 Hz, J₂=5.0 Hz, 1H), 7.85-7.87 (d, J=8.8 Hz, 2H),7.92-7.94 (m, 1H), 8.59-8.60 (dd, J₁=5.0 Hz, J₂=1.0 Hz, 1H), 8.69-8.70(d, J=2.7 Hz, 1H).

Example 88 Synthesis ofN-{5-[4-(pyridine-3-yloxy)phenyl]-1,3,4-oxadiazol-2-yl}-4-(trifluoromethoxy)benzamide

5-[4-(Pyridin-3-yloxy)phenyl]-1,3,4-oxadiazol-2-amine hydrobromide salt(167.5 mg, 0.5 mmol) was suspended in 2 mL of anhydrous pyridine.4-Trifluoromethoxybenzoyl chloride (167.2 mg, 117 uL, 0.75 mmol) wasadded directly into the solution. The reaction mixture formed anorange-red solution with a small amount of precipitate. It was left tostir for 6 hours. Then it was diluted with ca. 1 mL of MeOH, filteredthrough 0.22 u syringe filter and purified by reverse phase preparativeHPLC using acetonitrile/water mixture containing 0.01% of TFA.Fractions, containing the product, were combined and partitioned betweenca. 40 mL of EtOAc and ca. 40 mL of saturated aqueous NaHCO3. The EtOAclayer was washed with brine, dried over anhydrous sodium sulfate andfiltered. Solvent was removed in vacuo to give the title product as ayellow solid (89.0 mg). Yield 40.2%.

ESI-MS: [M+H]⁺ 442.82. ¹H NMR (DMSO-d₆): δ 7.20-7.22 (d, J=8.8 Hz, 2H),7.44-7.46 (d, J=8.6 Hz, 2H), 7.48-7.51 (dd, J=8.4 Hz, J₂=4.6 Hz, 1H),7.59-7.62 (ddd, J₁=8.4 Hz, J₂=2.8 Hz, J₃=1.2 Hz, 1H), 7.94-7.96 (d,J=8.8 Hz, 2H), 8.17-8.19 (d, J=8.6 Hz, 2H), 8.45-8.46 (dd, J₁=4.6 Hz,J₂=1.2 Hz, 1H), 8.48-8.49 (d, J=2.8 Hz, 1H).

Example 89 Synthesis ofN-{5-[4-(pyridin-3-yloxy)phenyl]-1,3,4-oxadiazol-2-yl}-3-(trifluoromethyl)benzamide

5-[4-(Pyridin-3-yloxy)phenyl]-1,3,4-oxadiazol-2-amine hydrobromide salt(167.5 mg, 0.5 mmol) was suspended in 2 mL of anhydrous pyridine.3-Trifluoromethylbenzoyl chloride (156.4 mg, 0.75 mmol) was added to thesolution. The reaction mixture formed an orange-red solution with asmall amount of white precipitate. It was left to stir for 3 hours. Thenit was diluted with ca. 1 mL of MeOH, filtered through 0.22 u syringefilter and purified by reverse phase preparative HPLC usingacetonitrile/water mixture containing 0.01% of TFA. Fractions,containing the product, were combined and partitioned between ca. 40 mLof EtOAc and ca. 40 mL of saturated aqueous NaHCO3. The EtOAc layer waswashed with brine, dried over anhydrous sodium sulfate and filtered.Solvent was removed in vacuo to give the title product as a yellow solid(50.0 mg). Yield 23.4%.

ESI-MS: [M+H]⁺ 426.94. ¹H NMR (DMSO-d₆): δ 7.21-7.23 (d, J=8.8 Hz, 2H),7.49-7.51 (dd, J₁=8.4 Hz, J₂=4.6 Hz, 1H), 7.60-7.62 (ddd, J₁=8.4 Hz,J₂=2.8 Hz, J₃=1.2 Hz, 1H), 7.75-7.76 (t, J=7.7 Hz, 1H), 7.95-7.98 (m,3H), 8.33-8.34 (d, J=7.7 Hz, 1H), 8.39 (s, 1H), 8.46-8.47 (dd, J₁=4.6Hz, J₂=1.2 Hz, 1H), 8.48-8.49 (d, J=2.8 Hz, 1H).

Example 90 Synthesis of4-Bromo-N-{5-[4-(pyridin-3-yloxy)phenyl]-1,3,4-oxadiazol-2-yl}-benzamide

5-[4-(Pyridin-3-yloxy)phenyl]-1,3,4-oxadiazol-2-amine hydrobromide salt(167.5 mg, 0.5 mmol) was suspended in 2 mL of anhydrous pyridine.4-Bromobenzoyl chloride (164.6 mg, 0.75 mmol) was added to the solution.The reaction mixture formed an orange-red solution with a small amountof white precipitate. It was left to stir for 3 hours. Then it wasdiluted with ca. 1 mL of MeOH, filtered through 0.22 u syringe filterand purified by reverse phase preparative HPLC using acetonitrile/watermixture containing 0.01% of TFA. Fractions, containing the product, werecombined and partitioned between ca. 40 mL of EtOAc and ca. 40 mL ofsaturated aqueous NaHCO3. The EtOAc layer was washed with brine, driedover anhydrous sodium sulfate and filtered. Solvent was removed in vacuoto give the title product as a yellow solid (46.5 mg). Yield 21.2%.

ESI-MS: [M+H]⁺ 438.84. ¹H NMR (DMSO-d₆): δ 7.20-7.22 (d, J=8.8 Hz, 2H),7.49-7.51 (dd, J₁=8.4 Hz, J₂=4.6 Hz, 1H), 7.60-7.62 (ddd, J=8.4 Hz,J₂=2.8 Hz, J₃=1.2 Hz, 1H), 7.68-7.70 (d, J=8.4 Hz, 2H), 7.95-7.97 (d,J=8.8 Hz, 2H), 8.00-8.02 (d, J=8.4 Hz, 2H), 8.45-8.46 (dd, J₁=4.6 Hz,J₂=1.2 Hz, 1H), 8.48-8.49 (d, J=2.8 Hz, 1H).

Example 91 Synthesis ofN-{5-[4-(pyridin-3-yloxy)phenyl]-1,3,4-oxadiazol-2-yl}-3-(trifluoromethoxy)-benzamide

5-[4-(Pyridin-3-yloxy)phenyl]-1,3,4-oxadiazol-2-amine hydrobromide salt(167.5 mg, 0.5 mmol) was suspended in 2 mL of anhydrous pyridine. Neat3-trifluoromethoxybenzoyl chloride (117 uL) was added directly into thesolution. The reaction mixture formed an orange-red solution with asmall amount of white precipitate. It was left to stir for 3 hours. Thenit was diluted with ca. 1 mL of MeOH, filtered through 0.22 u syringefilter and purified by reverse phase preparative HPLC usingacetonitrile/water mixture containing 0.01% of TFA. Fractions,containing the product, were combined and partitioned between ca. 40 mLof EtOAc and ca. 40 mL of saturated aqueous NaHCO3. The EtOAc layer waswashed with brine, dried over anhydrous sodium sulfate and filtered.Solvent was removed in vacuo to give the title product as a yellow solid(38.8 mg). Yield 17.5%.

ESI-MS: [M+H]⁺ 442.85. ¹H NMR (DMSO-d₆): δ 7.22-7.24 (d, J=8.8 Hz, 2H),7.49-7.51 (dd, J₁=8.4 Hz, J₂=4.6 Hz, 1H), 7.60-7.62 (ddd, J₁=8.4 Hz,J₂=2.8 Hz, J₃=1.2 Hz, 1H), 7.61-7.63 (m, 1H), 7.68-7.71 (t, J=7.7 Hz,1H), 7.97-7.98 (m, 3H), 8.08-8.10 (d, J=7.7 Hz, 1H), 8.46-8.47 (dd,J₁=4.6 Hz, J₂=1.2 Hz, 1H), 8.48-8.49 (d, J=2.8 Hz, 1H).

Example 92 Synthesis of4-methoxy-N-{5-[4-(pyridin-3-yloxy)phenyl]-1,3,4-oxadiazol-2-yl}-3-(trifluoromethyl)-benzamide

5-[4-(Pyridin-3-yloxy)phenyl]-1,3,4-oxadiazol-2-amine hydrobromide salt(167.5 mg, 0.5 mmol) was suspended in 2 mL of anhydrous pyridine. Neat4-methoxy-3-trifluoromethylbenzoyl chloride (179.0 mg, 0.75 mmol) wasadded directly into the solution. The reaction mixture formed anorange-red solution with a small amount of white precipitate. It wasleft to stir for 3 hours. Then it was diluted with ca. 1 mL of MeOH,filtered through 0.22 u syringe filter and purified by reverse phasepreparative HPLC using acetonitrile/water mixture containing 0.01% ofTFA. Fractions, containing the product, were combined and partitionedbetween ca. 40 mL of EtOAc and ca. 40 mL of saturated aqueous NaHCO₃.The EtOAc layer was washed with brine, dried over anhydrous sodiumsulfate and filtered. Solvent was removed in vacuo to give the titleproduct as a yellow solid (37.0 mg). Yield 16.2%.

ESI-MS: [M+H]⁺ 456.85. ¹H NMR (DMSO-d₆): δ 3.97 (s, 3H), 7.21-7.23 (d,J=8.8 Hz, 2H), 7.38-7.40 (d, J=9.1 Hz, 1H), 7.49-7.51 (dd, J₁=8.4 Hz,J₂=4.6 Hz, 1H), 7.60-7.62 (ddd, J₁=8.4 Hz, J₂=2.8 Hz, J₃=1.2 Hz, 1H),7.95-7.97 (d, J=8.8 Hz, 2H), 8.34-8.35 (m, 2H), 8.46-8.47 (dd, J₁=4.6Hz, J₂=1.2 Hz, 1H), 8.48-8.49 (d, J=2.8 Hz, 1H).

Example 93 Synthesis of2,2-Difluoro-N-{5-[4-(pyridin-3-yloxy)phenyl]-1,3,4-oxadiazol-2-yl}-1,3-benzodioxole-5-carboxamide

5-[4-(Pyridin-3-yloxy)phenyl]-1,3,4-oxadiazol-2-amine hydrobromide salt(167.5 mg, 0.5 mmol) was suspended in 2 mL of anhydrous pyridine. Neat2,2-difluoro-1,3-benzodioxole-5-carbonyl chloride (165.4 mg, 0.75 mmol)was added directly into the solution. The reaction mixture formed anorange-red solution with a small amount of white precipitate. It wasleft to stir for 3 hours. Then it was diluted with ca. 1 mL of MeOH,filtered through 0.22 u syringe filter and purified by reverse phasepreparative HPLC using acetonitrile/water mixture containing 0.01% ofTFA. Fractions, containing the product, were combined and partitionedbetween ca. 40 mL of EtOAc and ca. 40 mL of saturated aqueous NaHCO₃.The EtOAc layer was washed with brine, dried over anhydrous sodiumsulfate and filtered. Solvent was removed in vacuo to give the titleproduct as a yellow solid (27.0 mg). Yield 12.3%.

ESI-MS: [M+H]⁺ 456.85. ¹H NMR (DMSO-d₆): δ 7.19-7.21 (d, J=8.8 Hz, 2H),7.47-7.51 (m, 2H), 7.58-7.60 (ddd, J₁=8.4 Hz, J₂=2.8 Hz, J₃=1.2 Hz, 1H),7.93-7.95 (d, J=8.8 Hz, 2H), 7.99-8.01 (m, 2H), 8.45-8.46 (dd, J₁=4.6Hz, J₂=1.2 Hz, 1H), 8.48-8.49 (d, J=2.8 Hz, 1H).

Example 94 Synthesis of3-Chloro-2-fluoro-N-{5-[4-pyridin-3-yloxy)phenyl]-1,3,4-oxadiazol-2-yl}-5-trifluoromethyl)-benzamide

5-[4-(Pyridin-3-yloxy)phenyl]-1,3,4-oxadiazol-2-amine hydrobromide salt(167.5 mg, 0.5 mmol) was suspended in 2 mL of anhydrous pyridine. Neat3-chloro-2-fluoro-5-trifluoromethylbenzoyl chloride (250 uL) was addeddirectly into the solution. The reaction mixture formed a red solutionwith a small amount of white precipitate. It was left to stir for 18hours. Then it was diluted with ca. 1 mL of MeOH, filtered through 0.22u syringe filter and purified by reverse phase preparative HPLC usingacetonitrile/water mixture containing 0.01% of TFA. Fractions,containing the product, were combined and partitioned between ca. 40 mLof EtOAc and ca. 40 mL of saturated aqueous NaHCO₃. The EtOAc layer waswashed with brine, dried over anhydrous sodium sulfate and filtered.Solvent was removed in vacuo to give the title product as a yellow solid(83.4 mg). Yield 34.8%.

ESI-MS: [M+H]⁺ 480.71. ¹H NMR (DMSO-d₆): δ 7.18-7.20 (d, J=8.8 Hz, 2H),7.47-7.50 (dd, J₁=8.4 Hz, J₂=4.6 Hz, 1H), 7.58-7.60 (ddd, J₁=8.4 Hz,J₂=2.8 Hz, J₃=1.2 Hz, 1H), 7.92-7.94 (d, J=8.8 Hz, 2H), 8.13-8.16 (m,2H), 8.44-8.45 (dd, J=4.6 Hz, J₂=1.2 Hz, 1H), 8.47-8.48 (d, J=2.8 Hz,1H).

Example 95 Synthesis of4-Fluoro-N-{5-[4-(pyridin-3-yloxy)phenyl]-1,3,4-oxadiazol-2-yl}-3-(trifluoromethyl)-benzamide

5-[4-(Pyridin-3-yloxy)phenyl]-1,3,4-oxadiazol-2-amine hydrobromide salt(167.5 mg, 0.5 mmol) was suspended in 2 mL of anhydrous pyridine. Neat4-fluoro-3-trifluoromethylbenzoyl chloride (200 uL) was added directlyinto the solution. It was left to stir for 18 hours. The reactionmixture formed a red solution with a yellow precipitate. Then it wasdiluted with ca. 1 mL of MeOH, filtered through 0.22 u syringe filterand purified by reverse phase preparative HPLC using acetonitrile/watermixture containing 0.01% of TFA. Fractions, containing the product, werecombined and partitioned between ca. 40 mL of EtOAc and ca. 40 mL ofsaturated aqueous NaHCO3. The EtOAc layer was washed with brine, driedover anhydrous sodium sulfate and filtered. Solvent was removed in vacuoto give the title product as a yellow solid (105.1 mg). Yield 47.3%.

ESI-MS: [M+H]⁺ 444.79. ¹H NMR (DMSO-d₆): δ 7.18-7.20 (d, J=8.8 Hz, 2H),7.47-7.50 (dd, J₁=8.4 Hz, J₂=4.6 Hz, 1H), 7.53-7.60 (m, 2H), 7.93-7.94(d, J=8.8 Hz, 2H), 8.40-8.43 (m, 1H), 8.43-8.45 (dd, J₁=4.6 Hz, J₂=1.2Hz, 1H), 8.47-8.48 (d, J=2.8 Hz, 1H).

Example 96 Synthesis ofN-{5-[4-(pyridin-3-yloxy)phenyl]-1,3,4-oxadiazol-2-yl}-2-(trifluoromethoxy)-benzamide

5-[4-(Pyridin-3-yloxy)phenyl]-1,3,4-oxadiazol-2-amine hydrobromide salt(167.5 mg, 0.5 mmol) was suspended in 2 mL of anhydrous pyridine. Neat2-trifluoromethoxybenzoyl chloride (100 uL) was added directly into thesolution. It was left to stir for 18 hours. The reaction mixture formedan orange solution with a yellow precipitate. Then it was diluted withca. 1 mL of MeOH, filtered through 0.22 u syringe filter and purified byreverse phase preparative HPLC using acetonitrile/water mixturecontaining 0.01% of TFA. Fractions, containing the product, werecombined and partitioned between ca. 40 mL of EtOAc and ca. 40 mL ofsaturated aqueous NaHCO3. The EtOAc layer was washed with brine, driedover anhydrous sodium sulfate and filtered. Solvent was removed in vacuoto give the title product as a bright-yellow solid (20.2 mg). Yield9.1%.

ESI-MS: [M+H]⁺ 443.04. ¹H NMR (DMSO-d₆): δ 7.22-7.24 (d, J=8.8 Hz, 2H),7.48-7.54 (m, 4H), 7.60-7.63 (ddd, J₁=8.4 Hz, J₂=2.8 Hz, J₃=1.2 Hz, 1H),7.69-7.71 (t, J=7.8 Hz, 1H), 7.80-7.81 (d, J=7.5 Hz, 1H), 7.94-7.96 (d,J=8.8 Hz, 2H), 8.47-8.49 (m, 2H).

Example 97 Synthesis ofN-amino-N′-(4-chloro-3-trifluoromethyl-phenyl-guanidine hydroiodide

A mixture of 2.54 g 4-chloro-3-trifluoromethyl-phenylthiourea, 0.62 mlof iodomethane in 50 mL of anhydrous EtOH was refluxed for 1 hr to give1-[4-chloro-3-(trifluoromethyl)phenyl]-S-methylisothiourea hydroiodide.Then it was cooled down to ambient temperature and treated with 0.35 gof 98% hydrazine, heated gently with stirring until the initial vigorousevolution of MeSH subsided and then refluxed for additional 1 hour.

Example 98 Synthesis of4-(3-{[4-chloro-3-(trifluoromethyl)phenyl]amino}-1,2,4-triazin-5-yl)phenol

N-amino-N′-[4-chloro-3-(trifluoromethyl)phenyl]-guanidine hydroiodidecan be reacted with about 1.0-1.5 equivalents of4-hydroxy-phenylglyoxale in 1:1 mixture of methanol/water to yield thetitle product. The product can be isolated by precipitation or byextraction with a number of solvents, such as ethyl acetate, methylenechloride or diethyl ether or by silica gel column chromatography.

Example 99 Synthesis of4-[4-(3-{[4-chloro-3-(trifluoromethyl)phenyl]amino}-[1,2,4]triazin-5-yl)-phenoxy]-N-methylpyridine-2-carboxamide

7.23 g (19.73 mmol) of4-(3-{[4-chloro-3-(trifluoromethyl)phenyl]amino}-1,2,4-triazin-5-yl)phenolcan be dissolved in 80 mL of anhydrous DMF under argon atmosphere. 2.44g (21.71 mmol, 1.1 equivalent) of solid potassium tert-butoxide can beadded to the solution. The resulting mixture can be heated to about 100°C. and stirred at that temperature for 15 min. Then a solution of 3.7 g(21.71 mmol, 1.1 equivalent) of 4-chloro-pyridine-2-carboxylic acidmethylamide in 10 mL of anhydrous DMF can be added, followed by 3.28 g(23.68 mmol, 1.2 equivalent) of anhydrous K₂CO₃. The reaction mixturecan be heated at 140° C. for 30 hrs. The progress of the reaction can bemonitored by LC/MS. Then it can be allowed to cool down to ambienttemperature. The resulting mixture can be poured into 500 mL of waterand 100 mL of ethyl acetate. The aqueous layer can be extracted with anumber of solvents, such as ethyl acetate, methylene chloride or ether.The combined extracts can be washed 3 times with 100 mL of water, thenwith brine and dried over anhydrous sodium sulfate. Solvent can beremoved in vacuum to yield crude4-[4-(3-phenylamino-[1,2,4]triazin-6-yl)-phenoxy]-pyridine-2-carboxylicacid methylamide. The product can be than purified using silica gelcolumn chromatography. Those having ordinary skill in the art candetermine which solvent system can be used as an eluents in thechromatographic purification.

Example 100 Synthesis of 4-(3-ethylsulfanyl-[1,2,4]-triazin-5-yl-phenol

1.0-1.5 equivalent of 4-hydroxyphenylglyoxale can be reacted with1-amino-S-ethylisothiourea hydrobromide in 1:1 mixture of methanol/waterwith 1.0-2.0 equivalent of K₂CO₃ to yield4-(3-ethylsulfanyl-[1,2,4]triazin-5-yl)-phenol. The product can beisolated by a number of methods known to one skilled in the art, such asprecipitation or by extraction with a number of solvents, such as ethylacetate, methylene chloride or diethyl ether or by silica gel columnchromatography.

Example 101 Synthesis of 4-(3-amino-[1,2,4]-triazin-5-yl-phenol

1.0 equivalent of 4-(3-ethylsulfanyl-[1,2,4]triazin-6-yl)-phenol can bereacted with 1-5 equivalent of ammonia in dioxane to give4-(3-amino-[1,2,4]triazin-5-yl)-phenol. The product can be isolated by anumber of methods known to one skilled in the art, such as precipitationor by extraction with a number of solvents, such as ethyl acetate,methylene chloride or diethyl ether or by silica gel columnchromatography.

Example 102 Synthesis of4-[4-(3-amino-[1,2,4]triazin-5-yl)-phenoxy]-N-methylpyridine-2-carboxamide

3.71 g (19.73 mmol) of 4-(3-amino-[1,2,4]triazin-5-yl)-phenol can bedissolved in 80 mL of anhydrous DMF under argon atmosphere. 2.44 g(21.71 mmol, 1.1 equivalent) of solid potassium tert-butoxide can beadded to the solution. The resulting mixture can be heated to about 100°C. and stirred at that temperature for 15 min. Then a solution of 3.7 g(21.71 mmol, 1.1 equivalent) of 4-chloro-pyridine-2-carboxylic acidmethylamide in 10 mL of anhydrous DMF can be added, followed by 3.28 g(23.68 mmol, 1.2 eq.) of anhydrous K₂CO₃. The reaction mixture can beheated at 140° C. for 30 hrs. The progress of the reaction can bemonitored by LC/MS. Then it can be allowed to cool down to ambienttemperature. The resulting mixture can be poured into 500 mL of waterand 100 mL of ethyl acetate. The aqueous layer can be extracted with anumber of solvents, such as ethyl acetate, methylene chloride or ether.The combined extracts can be washed 3 times with 100 mL of water, thenwith brine and dried over anhydrous sodium sulfate. Solvent can beremoved in vacuum to give crude4-[4-(3-amino-[1,2,4]triazin-5-yl)-phenoxy]-N-methylpyridine-2-carboxamide.The product can be than purified using silica gel column chromatography.Those having ordinary skill in the art can determine which solventsystem can be used as an eluents in the chromatographic purification.

Example 103 Synthesis of4-[4-(3-{[4-chloro-3-(trifluoromethyl)benzoyl]amino}-[1,2,4]triazin-5-yl)-phenoxy]-N-methylpyridine-2-carboxamide

108.6 mg (0.337 mmol, 1.0 eq) of4-[4-(3-amino-[1,2,4]triazin-5-yl)-phenoxy]-N-methylpyridine-2-carboxamidecan be dissolved in 2 mL of anhydrous DMF with heating to about 100° C.45.4 mg (0.405 mmol, 1.2 equivalent) of solid t-BuOK can be added to thesolution, followed by 0.405 mmol (1.2 equivalent) of4-chloro-3-trifluoromethylbenzoyl chloride. It can be allowed to stir atambient temperature for 1-2 hours. The product can be isolated by anumber of methods known to those skilled in the art, such asprecipitation or by extraction with a number of solvents, such as ethylacetate, methylene chloride or diethyl ether or by silica gel columnchromatography, or by reverse-phase prep-HPLC chromatography.

Example 104 Synthesis of4-{4-[3-({[4-chloro-3-(trifluoromethyl)phenyl]sulfonyl}amino)-[1,2,4]triazin-5-yl)-phenoxy}-N-methylpyridine-2-carboxamide

108.6 mg (0.337 mmol, 1.0 equivalent) of4-[4-(3-amino-[1,2,4]triazin-5-yl)-phenoxy]-N-methylpyridine-2-carboxamidecan be dissolved in 2 mL of anhydrous pyridine with heating to about100° C. 0.405 mmol (1.2 equivalent) of4-chloro-3-trifluoromethyl-benzenesulfonyl chloride can be added. Thereaction mixture can be allowed to stir at ambient temperature for 1-2hours. The product can be isolated by a number of methods known to oneskilled in the art, such as precipitation or by extraction with a numberof solvents, such as ethyl acetate, methylene chloride or diethyl etheror by silica gel column chromatography, or by reverse-phase preparativeHPLC.

Example 105 Testing of Inhibition of MAPK Pathway in Cellular Assay

Some compounds described by the general structure (B) were tested forinhibition of MAPK pathway in cellular assay. Western Blot: Earlypassage primary human umbilical vein endothelial cells (HUVECs) weremaintained in EGM-2 containing SingleQuots (Cambrex, East Rutherford,N.J.), 10% FBS, 10 mM HEPES, and 50 μg/ml gentamicin. Prior to treatmentof the cells with inhibitor, the HUVECs were starved for 18 h byreplacing serum-containing complete media with serum-free andSingleQuot-free media. The starved cells were pre-treated withinhibitors for 60 min at various concentrations (0-20 μM). Next theHUVECs were treated with 50 ng/ml VEGF or FGF (Peprotech, Rocky Hill,N.J.) for 6 min and the cells were immediately washed with ice-cold PBS.Cells were lysed with ice-cold RIPA buffer containing 100 mM Tris pH7.5, 150 mM NaCl, 1 mM EDTA, 1% deoxycholate, 1% Triton X-100, 0.1% SDS,2 mM PMSF, one Complete-Mini protease inhibitor tablet (Roche,Indianapolis, Ind.; 1 tablet/7 ml of lysis buffer) and the phophataseinhibitors NaF (500 mM) and orthovanadate (1 mM). The cells were scrapedand lysates transferred and centrifuged at 15,000 g for 10 main.Supernatants were transferred to new tubes and protein concentration wasquantitated using the BCA protein reagent (Pierce, Rockford, Ill.). Celllysates containing 20 μg of total protein were separated by 10%SDS-PAGE, transferred to nitrocellulose, and blocked in 5% milk in TBST.Anti phospho-ERK Thr 202/Tyr 204 (Cell Signaling, Beverly, Mass.),anti-phospho-MEK Ser217/221 (Cell Signaling), and c-Raf (BD BiosciencesPharmingen, San Diego, Calif.) used as primary antibodies were detectedwith horseradish peroxidase-conjugated goat anti-mouse or rabbitsecondary antibodies and bands were visualized using the SuperSignalWest Pico chemiluminescence reagent system (Pierce) and Kodak X-ray film(Rochester, N.Y.).

Bay 43-9006 (Raf/FGF inhibitor) showed reduction of expression of p-MEKand p-ERK with IC50 between 200 and 300 nM when tested in this assay.U0126 (MEK inhibitor) showed reduction in p-Erk levels with IC₅₀ between200 and 300 nM, while p-MEK levels were unaffected. The results areshown in Table 1. As can be seen, compounds of the invention showedreduction in p-MEK and p-ERK levels with IC₅₀ between 400 nM and 20 μM.

Example 106 Cell Viability Assay

Some compounds described by the general structure (B) were tested forcell viability. XTT assay: HUVECs were seeded at 10,000 cells/well of atissue culture treated 96-well plate treated with collagen type I andgrown overnight in the complete EGM-2 media as described above. Thefollowing morning, the inhibitors were serial diluted with DMSO andadded to the cells with a final DMSO concentration of 1%. After 72 hourscell viability was measured with an XTT assay (Sigma, St. Louis, Mo.).The cells were also photographed to compare morphological differences tothe XTT trends observed. Determination of the IC₅₀ values was performedwith quantitative software (Prism software package, GraphPad Software,San Diego Calif.). Several inhibitors blocked cell proliferation andinduced apoptosis at concentrations below 1 μM and experiments wererepeated three times to confirm the observations. The compounds of theinvention displayed IC₅₀ between 100 nM and 40 uM in this assay (Table2).

TABLE 2 Test Results for Examples 105 and 106 Inhibition of HUVEC cellExamples Western Blot prolifiration (IC50)4-{4-[5-(4-chloro-3-trifluoromethyl-phenylamino)-4H-[1,2,4]triazol-3-active at 10 uM 2.85 uM yl]-phenoxy}-pyridine-2-carboxylic acidmethylamide4-[4-[5-(4-trifluoromethoxy-phenyl)-4H-[1,2,4]triazol-3-ylamino]- notactive 2.2 uM phenoxy}-pyridine-2-carboxylic acid methylamide(4-chloro-3-trifluoromethyl-phenyl)-{5-[4-(pyridine-3-yloxy)-phenyl]-active at 5 uM 1.81 uM 4H-[1,2,4]triazol-3-yl}-amine4-{4-[5-(4-trifluoromethoxy-phenylamino)-4H-[1,2,4]triazol-3-yl]- notactive >40 uM phenoxy}-pyridine-2-carboxylic acid methylamide{5-[4-(pyridine-4-yloxy)-phenyl]-4H-[1,2,4]triazol-3-yl}-(4- notactive >40 uM trifluoromethoxy-phenyl)amine6-{4-[5-(4-trifluoromethoxy-phenylamino)-4H-[1,2,4]triazol-3-yl]- notactive >20 uM phenoxy}-pyridazin-3-ylamine6-{4-[5-(4-trifluoromethoxy-phenylamino)-4H-[1,2,4]triazol-3-yl]- notactive 6.0 uM phenoxy}-pyrimidine-2,4-diamine6-[4-({5-[4-(trifluoromethoxy)-phenyl-4H-1,2,4-triazol-3- not active6.58 uM yl}amino)phenoxy]pyrimidine-2,4-diamine6-{4-[5-(4-chloro-3-trifluoromethyl-phenylamino)-4H-[1,2,4]triazol-3-active at 5 uM 0.089 uM yl]-phenoxy}-pyrimidine-2,4-diamine6-{4-[5-(4-chloro-3-trifluoromethyl-phenylamino)-4H-[1,2,4]triazol-3-not active 1.79 uM yl]-phenoxy}-pyridazin-3-ylamine4-{3-[5-(4-chloro-3-trifluoromethyl-phenylamino)-4H-[1,2,4]triazol-3-not active >20 uM yl]-phenoxy}-pyridine-2-carboxylic acid methylamide6-{3-[5-(4-chloro-3-trifluoromethyl-phenylamino)-4H-[1,2,4]triazol-3-active at 5 uM 0.404 uM yl]-phenoxy}-pyrimidine-2,4-diamine(4-chloro-3-trifluoromethyl-phenyl)-{5-[3-(pyridin-3-yloxy)-phenyl]-active at 5 uM 1.79 uM 4H-[1,2,4]triazol-3-yl}-amine6-{3-[5-(4-chloro-3-trifluoromethyl-phenylamino)-4H-[1,2,4]triazol-3-not active >10 uM yl]-phenoxy}-pyridazin-3-ylamine6-[4-(5-{[4-Chloro-3-trifluoromethyl-phenyl]amino}-1,3,4-oxadiazol-2-active at 5 uM 1.57 uM yl)-phenoxy]-pyrimidine-2,4-diamineN-[4-chloro-3-(trifluoromethyl)phenyl]-5-[4-(pyridin-3-yloxy)phenyl]-active at 5 uM 1.72 uM 1,3,4-oxadiazol-2-amineN-[4-chloro-3-(trifluoromethyl)phenyl]-5-[4-(pyridin-4-yloxy)phenyl]-active at 5 uM 8.6 uM 1,3,4-oxadiazol-2-amineN-[4-chloro-3-(trifluoromethyl)phenyl]-5-[4-(pyrimidin-5- active at 5 uM9.65 uM yloxy)phenyl]-1,3,4-oxadiazol-2-amine4-[4-(5-{[4-chloro-3-(trifluoromethyl)phenyl]amino}-1,3,4-oxadiazol-2-active at 5 uM 5.6 uM yl)phenoxy]-N-methylpyridine-2-carboxamide6-[4-({5-[4-chloro-3-(trifluoromethyl)phenyl-4H-1,2,4-triazol-3- activeat 5 uM 9.57 uM yl}amino)phenoxy]pyrimidine-2,4-diamine5-[4-chloro-3-(trifluoromethyl)phenyl]-N-[4-(pyridine-4-yloxy)phenyl]-not active >40 uM 4H-1,2,4-triazol-3-amine5-[4-chloro-3-(trifluoromethyl)phenyl]-N-[4-(pyrimidin-5- active at 5uM >10 uM yloxy)phenyl]-4H-1,2,4-triazol-3-amine6-[4-({5-[4-chloro-3-(trifluoromethyl)phenyl]-4H-1,2,4-triazol-3- activeat 5 uM >40 uM yl}amino)phenoxy]pyridazin-3-amine4-[4-({5-[4-chloro-3-(trifluoromethyl)phenyl]-4H-1,2,4-triazol-3- notactive >40 uM yl}amino)phenoxy]-N-methylpyridine-2-carboxamide5-[4-chloro-3-(trifluoromethyl)phenyl]-N-[4-(pyridine-3-yloxy)phenyl]-active at 5 uM >40 uM 4H-1,2,4-triazol-3-amine6-[4-(5-{[3-(Trifluoromethyl)phenyl]amino}-1,3,4-oxadiazol-2- active at5 uM >10 uM yl)phenoxy]pyrimidine-2,4-diamine5-[4-(Pyrimidin-5-yloxy)phenyl]-N-[3-(trifluoromethyl)phenyl]-1,3,4-active at 5 uM >10 uM oxadiazol-2-amine6-(4-{5-[(4-chlorophenyl)amino]-1,3,4-oxadiazol-2-yl}phenoxy) active at5 uM 2.6 uM pyrimidine-2,4-diamine5-[4-(Pyrimidin-5-yloxy)phenyl]-N-[4-chloro-phenyl]-1,3,4-oxadiazol-2active at 5 uM >20 uM amine6-(4-{5-[(2-chloro-5-trifluoromethyl-phenyl)amino]-1,3,4-oxadiazol-2-not active ~20 uM yl}phenoxy) pyrimidine-2,4-diamine5-[4-(Pyrimidin-5-yloxy)phenyl]-N-[2-chloro-5-(trifluoromethyl)- notactive ~20 uM phenyl]-1,3,4-oxadiazol-2-amine6-(4-{5-[(3-chlorophenyl)amino]-1,3,4-oxadiazol-2-yl}phenoxy) active at5 uM ~20 uM pyrimidine-2,4-diamine5-[4-(Pyrimidin-5-yloxy)phenyl]-N-[2-chloro-phenyl]-1,3,4-oxadiazol-2-active at 5 uM ~20 uM amine5-[4-(Pyridin-3-yloxy)phenyl]-N-[4-chloro-phenyl]-1,3,4-oxadiazol-2-active at 10 uM 9.6 uM amine5-[4-(Pyridin-3-yloxy)phenyl]-N-[3-trifluoromethyl-phenyl]-1,3,4- activeat 10 uM >20 uM oxadiazol-2-amine5-[4-(Pyridin-3-yloxy)phenyl]-1,3,4-oxadiazol-2-amine not active at 10uM >40 uM N-{5-[4-(pyridin-3-yloxy)phenyl]-1,3,4-oxadiazol-2-yl}-4- notactive at 10 uM >40 uM (trifluoromethoxy)benzamideN-{5-[4-(pyridin-3-yloxy)phenyl]-1,3,4-oxadiazol-2-yl}-3- active at 10uM >40 uM (trifluoromethoxy)benzamide4-Bromo-N-{5-[4-(pyridin-3-yloxy)phenyl]-1,3,4-oxadiazol-2-yl}- notactive at 10 uM >40 uM benzamideN-{5-[4-(pyridin-3-yloxy)phenyl]-1,3,4-oxadiazol-2-yl}-3- not active at10 uM >40 uM (trifluoromethoxy)-benzamide4-Methoxy-N-{5-[4-(pyridin-3-yloxy)phenyl]-1,3,4-oxadiazol-2-yl}-3-active at 10 uM >40 uM (trifluoromethoxy)-benzamide2,2-Difluoro-N-{5-[4-(pyridin-3-yloxy)phenyl]-1,3,4-oxadiazol-2-yl}- notactive at 10 uM >40 uM 1,3-benzodioxole-5-carboxamide3-Chloro-2-fluoro-N-{5-[4-(pyridin-3-yloxy)phenyl]-1,3,4-oxadiazol-2-not active at 10 uM >40 uM yl}-5-(trifluoromethoxy)-benzamide4-Fluoro-N-{5-[4-(pyridin-3-yloxy)phenyl]-1,3,4-oxadiazol-2-yl}-3-active at 10 uM ~20 uM (trifluoromethoxy)-benzamideN-{5-[4-(pyridin-3-yloxy)phenyl]-1,3,4-oxadiazol-2-yl}-2- not active at10 uM ~20 uM (trifluoromethoxy)-benzamide

Although the invention has been described with reference to the aboveexamples, it will be understood that modifications and variations areencompassed within the spirit and scope of the invention. Accordingly,the invention is limited only by the following claims.

1. A compound having the structure (A) or an N-oxide, N,N′-dioxide,N,N′,N″-trioxide, or a pharmaceutically acceptable salt thereof:

wherein: Y is absent or is a moiety selected from a group consisting of:

R₁ is a substituent selected from a group consisting of an aryl, asubstituted aryl, a heterocycle, a heteroaryl, a substitutedheterocycle, and a substituted heteroaryl; R₂ is a substituent selectedfrom a group consisting of hydrogen, hydroxyl, C₁-C₆ alkyl, C₁-C₆alkylamino, halogen, C₁-C₆ alkoxy, —NO₂, —NH₂, and —C≡N; and R₃ is asubstituent selected from a group consisting of an aryl, a substitutedaryl, a heterocycle, a heteroaryl, a substituted heterocycle, and asubstituted heteroaryl.
 2. The compound of claim 1, wherein: R₁ isselected from a group consisting of C₆-C₁₂ aryl; C₃-C₁₂ heteroarylhaving 1-3 heteroatoms selected from N, S and O, substituted C₃-C₁₀cycloalkyl having 0-3 heteroatoms selected from N, S, and O, substitutedC₆-C₁₂ aryl, substituted C₃-C₁₂ heteroaryl having 1-3 heteroatomsselected from N, S and O, C₇-C₂₄ aralkyl; C₇-C₂₄ alkylaryl; substitutedC₇-C₂₄ aralkyl; and substituted C₇-C₂₄ alkaryl; R₃ is selected from agroup consisting of C₆-C₁₂ aryl; C₃-C₁₂ heteroaryl having 1-3heteroatoms selected from N, S and O, substituted C₃-C₁₀ cycloalkylhaving 0-3 heteroatoms selected from N, S, and O, substituted C₆-C₁₂aryl, substituted C₃-C₁₂ heteroaryl having 1-3 heteroatoms selected fromN, S and O, C₇-C₂₄ aralkyl; C₇-C₂₄ alkylaryl; substituted C₇-C₂₄aralkyl; and substituted C₇-C₂₄ alkaryl.
 3. The compound of claim 1,wherein the compound having the structure (A) is selected from the groupof compounds having the structures (A1), (A2), (A3), and (A4):


4. The compound of claim 3, wherein the compound has the structure (A1)and wherein: R₁ is selected from a group consisting of

R₃ is selected from a group consisting of:

and n is an integer selected from a group consisting of 0, 1, 2, and 3.5. The compound of claim 3, wherein the compound having the structure(A1) is selected from a group consisting of compounds having structures(1) and (2):


6. The compound of claim 3, wherein the compound having structure (A1)is selected from a group consisting of compounds having the structures(3), (4), (5) and (6):


7. The compound of claim 3, wherein the compound having structure (A1)is selected from a group consisting of compounds having the structures(7)-(16):


8. The compound of claim 7, wherein the compound has the formula:


9. The compound of claim 7, wherein the compound has the formula:


10. The compound of claim 7, wherein the compound has the formula:


11. The compound of claim 7, wherein the compound has the formula:


12. The compound of claim 7, wherein the compound has the formula:


13. The compound of claim 7, wherein the compound has the formula:


14. The compound of claim 7, wherein the compound has the formula:


15. The compound of claim 7, wherein the compound has the formula:


16. The compound of claim 7, wherein the compound has the formula:


17. The compound of claim 7, wherein the compound has the formula:


18. A pharmaceutical composition comprising a compound set forth inclaim 1 and a pharmaceutically acceptable carrier.
 19. A pharmaceuticalcomposition comprising a compound as set forth in claim 3, wherein thecompound is of Structure (A1), and a pharmaceutically acceptablecarrier.
 20. A pharmaceutical composition comprising a compound as setforth in claim 3, wherein the compound is of Structure (A2), and apharmaceutically acceptable carrier.
 21. A pharmaceutical compositioncomprising a compound as set forth in claim 3, wherein the compound isof Structure (A3), and a pharmaceutically acceptable carrier.
 22. Apharmaceutical composition comprising a compound as set forth in claim3, wherein the compound is of Structure (A4), and a pharmaceuticallyacceptable carrier.